Limited pulmonary resections for congenital cystic adenomatoid malformation of the lung. Congenital cystic adenomatoid malformations (CAMs) are rare lung lesions characterized by the presence of a multicystic mass of pulmonary tissue. To clarify the surgical management of CAM of the lung, we reviewed our institutional experience with 34 patients with histologically documented CAM. Symptoms developed in the first 2 days of life in 20 patients; 14 patients developed symptoms a median of 69 days (range, 22 days to 15 years) after birth. A diagnosis of CAM was confirmed at thoracotomy in all patients. Four patients were treated with a limited pulmonary resection for disease confined to one lobe. Seven patients underwent a composite resection involving a lobectomy plus limited resection of the second lobe. One patient underwent pneumonectomy for a severely malformed lung. The remainder of the patients were treated with anatomic lobectomy. The only recurrence was a patient who had a partial cystectomy performed at the initial operation. We conclude that in the appropriate malformation subtype, limited pulmonary reactions can preserve lung tissue and may prevent subsequent complications of CAM.


Prenatal ultrasonography and early surgery for congenital cystic disease of the lung. With the recent advent of prenatal ultrasound as a routine screening procedure, diagnosis of congenital cystic lung disease has been made in utero, raising the possibility of elective surgery for these lesions early in infancy before the patient develops respiratory distress or potentially life-threatening infection. From 1979 to 1989 six cases of congenital lung cyst were diagnosed in utero by prenatal ultrasound and followed during pregnancy. Two of the six were not confirmed after birth because the mothers preferred an abortion. The remaining four cases were studied periodically during gestation by ultrasonography. At birth, the first infant developed respiratory distress and underwent urgent left upper lobectomy and left lower segmentectomy at age 18 hours. The other three underwent elective lobectomy at age 10 days, 3 months, and 7 months, respectively. The fourth infant had a normal chest x-ray and ultrasound at birth, and the congenital cysts were confirmed by computed tomography scan. The pathological diagnosis in all four cases was cystic adenomatoid malformation. In two cases, intraoperative measurement of pulmonary function demonstrated significant improvement after resection of the affected lobe. We conclude that congenital lung cysts can be accurately diagnosed by prenatal ultrasound "screening" as early as 18 to 24 weeks' gestation. Advantages of early diagnosis include the option of moving the mother and unborn child to a high-risk obstetrical center for urgent operation on the newborn infant if necessary. Otherwise, once the diagnosis is confirmed, surgical correction can be performed electively and safely before respiratory distress or pulmonary infection complicates the infant's growth and development.


Surgery for achalasia of the cardia in children: the Dor-Gavriliu procedure. The Dor-Gavriliu procedure was utilized in six children with achalasia of the cardia from 1983 to 1991. Four boys and two girls (mean age, 10.1 years; range, 3 months to 16 years) presented with symptoms of weight loss (83%), emesis (83%), dysphagia (67%), recurrent respiratory infections (67%), and nocturnal regurgitation (33%). The diagnosis of achalasia was established by barium swallow in all patients; esophageal manometry was used in four patients to confirm the diagnosis. Follow-up ranged from 10 months to 8 years (mean, 3.5 years). Four neurologically normal patients had excellent results with complete resolution of their preoperative symptoms. Two neurologically impaired children, both afflicted with Down's syndrome, had less than excellent results. One moderately impaired child had a good result (required three postoperative bougie dilations over 8 years without demonstration of gastroesophageal reflux); the second, more severely impaired child, had only a fair result (persistent failure to thrive with the development of grade II gastroesophageal reflux). The Dor-Gavriliu procedure uses a transabdominal, anterior esophageal myotomy with incorporation of an effective, nonobstructing, antireflux mechanism that should prevent myotomy reapposition.


Sugiura procedure in the surgical treatment of bleeding esophageal varices in children: long-term results. Recurrent bleeding, the high occurrence of encephalopathy, and the impairment of hepatic function in the successful cases of portasystemic shunts have led to increasing dissatisfaction with these procedures in recent years. Between March 1974 and November 1990 we have operated on 15 children for bleeding esophageal varices using the Sugiura procedure (esophageal transection with paraesophagogastric devascularization). In two cases the entire procedure was performed via the thoracic approach. The spleen was left in place in five cases. We have had no mortality. Operative complications included bleeding in the early postoperative period in two children and partial leakage from the esophageal suture in two others. Follow-up was from 4 years 3 months to 16 years 8 months with an average of 10 years 4 months. Long-term results have been gratifying in 12 patients (80%) with disappearance of the varices and no evidence of recurrent bleeding. Three children (20% of the patients) had recurrent bleeding 4 1/2 years, 4 years 2 months, and 2 1/2 years after the surgical procedure. In all cases there was no evidence of esophageal stenosis, gastroesophageal reflux, or hiatal hernia, nor signs of encephalopathy nor impairment of hepatic function. In our opinion the Sugiura procedure is a valid procedure in the surgical treatment of esophageal varices bleeding in the pediatric age with a high rate of success (80%) and without late complications.


Postobstructive enteropathy in infants with transient enterostomy: its consequences on the upper small intestinal functions. Repeated or prolonged organic obstruction of the small intestine in the neonatal period can lead to severe refeeding problems, despite a transient ostomy. These problems are thought to result from a postobstructive enteropathy (POE) of the apparently normal small intestine segment above the obstruction. Ten infants with a POE, characterized by limited oral caloric and carbohydrate intakes and increased ostomy effluent, were compared with 8 controls with an enterostomy and a normal postoperative refeeding pattern. There was no statistical difference in the histomorphometric appearance of the mucosa or its digestive or absorptive capacity (brush-border hydrolases, glucose transport) between the two groups. The effluent and duodenal floras of the two groups were similar. However, all POE patients showed significant abnormal peristalsis characterized by barium and carmin transit times. This suggests that repeated or prolonged obstruction in the neonatal period could lead to a POE, caused by chronic motricity abnormalities of the small intestine above the obstruction. Although this POE is more frequent after small bowel atresia, it may also occur with other conditions causing prenatal and postnatal intestinal obstruction.


Nonischemic intussusception in childhood. The classical presentation of intussusception consisting of severe abdominal pain, bloody stool, and a palpable abdominal mass leads to the correct diagnosis in majority of the patients. However, an atypical presentation often results in a delayed diagnosis as is commonly seen in nonischemic intussusception. The nonischemic intussusception is a distinct clinical entity that is characterized by a long history of less severe symptoms commonly noticed in older children. The incidence of diarrhea in this group is higher than in the acute variety of intussusception. This variant of intussusception requires a high degree of suspicion for the diagnosis in atypical clinical presentation. The present study summarises our experience treating 31 such cases of nonischemic intussusception during a period of 25 years from 1966 to July 1990.


Familial intussusception.

Familial intussusception. We report a father and two sons who each suffered from recurrent acute ileocolic intussusception in childhood, suggesting that there may in some cases be a genetic predisposition to the condition. This might have an anatomical basis.


Sites of relapse in patients with neuroblastoma following bone marrow transplantation in relation to preparatory "debulking" treatments. Forty-one patients with high-risk neuroblastoma were treated between September 1977 and December 1987 at the Children's Hospital of Philadelphia with supralethal chemotherapy and total-body irradiation rescued by bone marrow transplantation. Twenty-six patients were treated following relapse and 15 were newly diagnosed. At the time of evaluation, January 1991, 11 of 41 patients (26.8%) remained in complete remission. Actuarial survival rates of patients transplanted following relapse were 0.35 and 0.31 at 2 and 5 years, respectively, and actuarial disease-free survival rates were 0.38 at 12 months and 0.27 at 24 months. The 2- and 5-year actuarial survival values for the patients with newly diagnosed disease were 0.53 and 0.25, respectively, and the 12- and 24-month disease-free survival rates were 0.47 and 0.27, respectively. There was no significant difference in survival between these groups. Twenty-nine of the 41 patients reviewed were available for analysis of the effect of local treatment. Thirteen had a combination of surgery and radiation (RT), 2 had surgery alone, 9 had RT alone, and in 5 patients no local treatment was given. The local relapse rate was 17%; it was 15% following surgery plus RT and 22% following RT alone. The failure rate combining local and distant relapse is 62% for surgery plus RT and 44% for RT alone. Although a local relapse rate of 17% is imperfect, it is a relatively small contribution to the overall relapse of 62%.


Cystic lesions of the fetal kidney: diagnosis and prediction of postnatal function by fetal urine biochemistry. Fetal urine was aspirated under ultrasound control from 21 large cystic renal masses in 18 pregnancies of 20 to 35 weeks gestation. None were associated with bladder or ureteric dilatation. At postnatal investigation, 12 kidneys were demonstrated to be hydronephrotic (5 with no or poor function) and 9 multicystic. Urinary concentrations of sodium (Na+), calcium (Ca++), and phosphate (PO4--) were significantly higher in the multicystic group than in the hydronephrotic, whereas urea and creatinine levels were lower. Determination of urinary PO4-- enabled differential diagnosis with no false-positive or false-negative cases. Among hydronephrotic kidneys, no biochemical parameter accurately predicted postnatal function, although creatinine was increased in all three nonfunctioning kidneys. In hydronephrotic kidneys, urinary Na+ concentration increased with advancing gestational age (r = .66; P < .02), suggesting that the duration of hydronephrosis has a negative effect on renal function.


Ectopic ureter with complete ureteric duplication in the female child. Twenty-two female children with complete ureteric duplication and ectopic ureter (two bilateral) were seen in a 15-year period. The most common clinical presentation was dribbling urinary incontinence with "normal" micturition. Other presentations included urinary infection and vaginal discharge. Four cases were diagnosed after antenatal recognition of ureterohydronephrosis. The clinical diagnosis was supported by various radiological investigations but ultrasonography (US) proved to be particularly reliable in diagnosing ectopic ureter. The most common sites of opening of the ectopic ureter were the urethral margin or the urethrovaginal septum, although in seven cases the site was not identified. Twenty-one kidneys were managed by upper pole heminephrectomy and three by ureteropyelostomy, removing as much of the ectopic ureter as possible via the renal approach. The distal ectopic ureter was removed via a separate suprapubic incision at the initial operation in four cases, and in two cases, delayed excision of the distal ectopic ureter was necessary. All surgical specimens were examined histologically and only two heminephrectomy specimens showed features of renal dysplasia. Dribbling urinary incontinence was cured in all cases, although in one patient the entire kidney was lost after heminephrectomy. Ectopic ureter should be suspected in girls with dribbling urinary incontinence. The diagnosis is best supported by US together with conventional radiology. The majority of cases can be managed by heminephrectomy, but when adequate function is demonstrated in the upper pole, ureteropyelostomy is recommended.


Neuropathic urinary incontinence in pediatric patients: management with artificial sphincter. From June 1982 to November 1990, 37 patients, aged 13 to 19 years (35 males and 2 females), were treated in our division for neuropathic urinary incontinence with an artificial sphincter. All patients were treated before sphincter implantation with drugs, transurethral sphincterotomy in boys, and bladder flap urethroplasty was carried out in females during the surgical procedure. The cuff was placed at the bladder neck in 33 cases, and in 4 cases at the urethral bulb. The sphincter was activated about 3 weeks after implantation and after 2 months in two patients with associated bladder augmentation. Operative and perioperative complications occurred in four cases during the initial phase of our experience. Later we had to perform 19 revisions on 14 patients due to mechanical and surgical failure (reoperation rate, 0.38). Thirty-three patients are presently dry (90% of successes). Postoperatively, normal upper urinary tract conditions were found in all patients except two.


Repair of posttraumatic stenosis of the urethra through a posterior sagittal approach. The authors present a report of four cases of children with posttraumatic posterior urethral stenosis who were surgically corrected through a posterior sagittal approach, with resection of the stenotic area and end-to-end anastomosis. In all cases whole colonic irrigation was used to prepare the bowel for surgery without the use of colostomy. The results were good in all cases and there were no complications. The anatomic bases of the surgical approach as well as the technical details of the procedure are analyzed, and the advantages are compared with other techniques used for the same purpose.


Vibro-acoustic stimulation does not alter the duration of high and low fetal heart rate variability episodes. To study the effect of vibro-acoustic stimulation on high and low fetal heart rate (FHR) variability episodes. Prospective control study, in 21 high risk pregnancies between 32 to 39 weeks gestation. FHR pattern was recorded with cardiotocograph (CTG). After 5 min low FHR variability a vibro-acoustic stimulator was activated in contact with or above the maternal abdomen (sham) on a random basis on two consecutive days, and CTG was continued to obtain a complete high and low FHR variability cycle. Complete high and low FHR variability cycles were obtained in 12 patients with mean recording time of 129.5 minutes (range 69-185 min). The mean baseline FHR increased by 32.5 +/- 9.4 bpm after contact vibro-acoustic stimulation, but remained unchanged after sham stimulation. Mean duration of high FHR variability was 56.6 +/- 32.9 min and 47.3 +/- 23.7 min, respectively, and mean duration of the consecutive low FHR variability episode was similar after contact and sham stimulation. The complete cycles were 71.1 +/- 34.7 min and 66.2 +/- 21.2 min, respectively. Vibro-acoustic stimulation induced an abrupt change from low to high FHR variability, but the complete cycle of high and low FHR variability was not altered, when compared with the corresponding cycle following sham stimulation on the same fetus.


Fetal cerebral haemodynamic adaptation: a progressive mechanism? Pulsed and color Doppler evaluation. The importance of studying, by Doppler ultrasound, the cerebral haemodynamics to monitor the fetal response to the hypoxia is well known, but there is not a general agreement about the anatomical landmarks for the middle cerebral artery. Seventy-one normal fetuses and fifteen IUGR fetuses were studied. The umbilical artery and the middle cerebral artery (MCA) were evaluated by color Doppler ultrasound. The well-known decrease of the pulsatility index from the umbilical artery was observed throughout pregnancy. This velocimetric pattern did not occur in the IUGR fetuses. Four IUGR fetuses showed the ARED (absent or reversed and diastolic flow) pattern. The MCA was evaluated at the origin (M1) and at the distal tract (M2). PI values from M1 and M2 decreased during the pregnancy. A significant difference was detected between M1 and M2 PI values from the 26th to the 37th week of gestation. The M1 brain sparing effect was detected in the IUGR fetuses. Two ARED fetuses, observed during labor, showed the M2 sparing effect. The different Doppler patterns found in M1 and M2 could be due to the functional differences existing between these tracts. As a matter of fact, M1 and M2 supply different parts of the fetal brain, which develop in different periods of fetal life. These findings, if ulteriorly confirmed, could offer new perspectives for the monitoring of high risk fetuses.


Early-diastolic blood flow velocity block in the umbilical artery and fetal cardiac valvular insufficiency. Case report. Absence of early-diastolic blood flow velocity was recorded from the umbilical artery of a fetus with severe myocardial and valvular insufficiency. The blood flow at late diastole was normal. This flow pattern may indicate the need for a detailed ultrasonic examination of the fetal heart.


Prediction of RDS by amniotic fluid analysis: a comparison of the prognostic value of traditional and recent methods. The determination of lecithin or even more the lecithin/sphingomyelin (L/S) ratio in amniotic fluid are both well established in the prediction of neonatal RDS. The immunological measurement of phosphatidylglycerol (PG) and the determination of the surfactant/albumin (S/A) ratio by fluorescence polarization (TDx FLMR) have recently been introduced for the detection of fetal lung maturity. In order to compare traditional versus recent methods L/S ratio and PG determination by one dimensional thin-layer chromatography, enzymatic analysis of lecithin, immunological determination of PG by Amniostat-FLMR and the fluorescence polarization of S/A-ratio by the TDx FLMR were all performed in 141 amniotic fluid samples of 122 patients. Only one out of 72 samples was false negative in the enzymatic lecithin determination (sensitivity 88%). All other methods have a sensitivity and a negative predictive value of 100%. The positive predictive values and the specificity varied between 22%-50% and 58%-87% respectively. The false positive rate, which is high for all methods, is lowest for the L/S ratio. This study demonstrates, that the recent methods are reasonable alternatives in all cases with a positive test. In clinical practice they have the advantage, that the result can be obtained in 15 minutes. If the test is predictive for lung immaturity the L/S ratio should be performed in addition to decrease the false positive rate before any clinical decision is made.


Short-term effect of magnesium sulfate infusion on renal function in preeclamptic women. Magnesium (Mg) plays an important role in cardiovascular homeostasis. A deficiency may be of importance for the etiology of disorders that have vasospasm in common. Mg administration can reduce the peripheral vascular resistance and thus enhance organ perfusion. We observed that Mg sulfate infusion could have a beneficial effect upon the serum urate concentration in preeclamptic women, presumably by affecting the renal function. A study comprising 10 preeclamptic women with a high serum urate level (413-788 umol/l) was carried out. Glomerular filtration rate (GFR) was measured by determination of iohexol clearance. 30 mmol Addex-Magnesium was then given i.v. during 12 h and a second GFR determination performed the next day. We had expected the GFR to increase, but to our surprise, it decreased (in mean, from 74.2 ml/[min x 1.73 m2] to 71.3; p < 0.05; Wilcoxon signed-ranks test). There were no significant changes of blood pressure, proteinuria, blood thrombocytes, transaminases, serum creatinine or serum urate. We conclude that in preeclamptic women with renal dysfunction, Mg infusion therapy had no favourable effect upon either blood pressure nor GFR in the short-term perspective.


Measurement of systolic blood pressure in the follow-up of low birth weight infants. Systolic blood pressure was measured using the Doppler technique in 149 infants who attended the neonatal follow-up clinic. One hundred and six of these infants were graduates of the neonatal intensive care unit and 81% were prematurely delivered. Most of the participants were examined only once and their postnatal age ranged between 2-191 weeks at the time the blood pressure was measured. During the first weeks of life the blood pressure in infants whose birth weight was > 2000 grams appeared to be higher than that of infants of lower birth weight infants (< 2000 grams), but the difference disappeared within the first months of life. Almost all measurements after 6 months of age were of lower birth weight infants (< 2000 grams) and their systolic blood pressure level of 101.1 +/- 6.4 mm Hg (mean +/- S.D.; n = 38) appeared to remain unchanged in later infancy. Using analysis of variance, four perinatal factors were found to be associated by analysis of variance with elevated blood pressure values recorded in the clinic. These were instrumental vaginal delivery, prolonged ventilatory support, presence of patent ductus arteriosus and high blood pressure during the neonatal period. However, when multiple regression analysis was applied, only neonatal hypertension was significantly associated with subsequent elevated values measured in the clinic.


Analysis of the tcPO2 response to blood interruption in infants and adults. The transcutaneous PO2 (tcPO2) response to blood interruption (BIS test) was measured in 6 healthy adults and 28 infants, including premature infants. The response was similar in both infants and adults with a time lag at the beginning of blood interruption, a subsequent linear decrease, a time lag at the end of blood interruption, and an exponential recovery to the final steady state. The linear decrease of tcPO2 in the BIS test had a correlation coefficient of more than 0.98 in all subjects and indicated the cutaneous oxygen consumption rate independent of the oxygen pressure. There was a significant difference in oxygen consumption according to birth weight in the infants (< 2 kg, 2-3 kg, and > 3 kg). The recovery time constant for infants was 13 seconds which was about 1.4 times faster than that for adults, and it increased significantly along with a higher birth weight. The time lag at blood interruption was longer than that at blood recovery. The time lag for infants was significantly shorter than that for adults, and in infants there was a significant difference between the group with a birth weight < 2 kg and the others (p < 0.005). All of the parameters of the BIS test suggested the physiological immaturity of premature infants.


Raised plasma hypoxanthine levels as a prognostic sign in preterm babies with respiratory distress syndrome treated with natural surfactant. Plasma hypoxanthine concentration was measured in twelve preterm babies with respiratory distress syndrome (RDS) treated with 200 mg/kg of a porcine surfactant (Curosurf). Five of the babies died within one week and seven survived the neonatal period. Surviving babies had no significant changes in plasma hypoxanthine concentration throughout a one hour study period following the administration of surfactant. By contrast, in nonsurvivors the mean plasma hypoxanthine concentrations increased from 6.8 mumol/l before surfactant administration to 14.2 mumol/l 15 minutes after surfactant treatment. Survivors had a mean maximal increase in plasma hypoxanthine of 1.9 mumol/l 15-30 min factor surfactant treatment compared with 9.4 mumol/l in nonsurvivors (p < 0.05). The babies who developed intracranial hemorrhage had significantly higher maximal plasma hypoxanthine increase (mean 9.6 mumol/l) compared with babies who did not develop intracranial hemorrhage (mean 1.1 mumol/l) (p < 0.01). The combination of high PaO2 and high hypoxanthine concentration could lead to an increased production of oxygen radicals which might be harmful. We conclude that plasma hypoxanthine concentration may serve as an indicator of the prognosis in preterm babies treated with natural surfactant. Further, it seems important to reduce oxygen supplementation as soon as surfactant is given to possibly limit oxygen radical production.


Uteroplacental diastolic notching in 510 uneventful pregnancies. In a collective of 510 patients with uneventful pregnancies and deliveries the rate of waveform notching in the uteroplacental flow velocity pattern was examined. Notching appeared to be significantly more frequent in the uterine than arcuate artery. It was unilateral and mainly affected placenta-contralateral sites. Bilateral notching or notching on placental site were the exception. In both flow patterns notching was seen to increase as pregnancy progressed and was slightly higher in the third compared to the second trimester.


Fetal hydrocephalus associated with maternal neurofibromatosis. Neurofibromatosis (NF), an autosomal dominant inherited disorder affecting multiple organ systems, is rare among pregnant women. NF in pregnancy has been reported to be complicated by maternal hypertension, fetal intrauterine growth retardation and fetal wastage. A case of pregnancy associated with neurofibromatosis, complicated by pregnancy aggravated chronic hypertension, asymmetric intrauterine growth retardation and fetal hydrocephalus is described. Possible interrelations between those pathologies are discussed.


Principles and pitfalls of clinical trials design. Periodontitis and gingivitis trials can involve many complex experimental designs. The selection of a specific design and the details of the protocol can influence the magnitude of any effect observed, the generalizability of the results, and the clinical significance of the findings. The purpose of this paper is to review selected aspects of clinical trials including the overall clinical experimental design, controls, outcomes, sample size, and patient selection. Particular emphasis will be placed on periodontitis trials.


Differential diagnosis in clinical trials of therapy for periodontitis. For purposes of clinical trials of therapies for periodontitis, it is recommended that population samples be identified as adult periodontitis (systemically modified or not systemically modified), early onset periodontitis (localized or juvenile, generalized or rapidly progressive, or associated with systemic disease), or necrotizing ulcerative periodontitis, avoiding overlapping criteria. Population samples of adult or early onset periodontitis modified by or associated with systemic conditions should be used in clinical trials only when the intent is to study effect on the specifically modified condition. Objective identification of the modifying systemic condition should be required for all subjects in such trials. Population samples should be homogeneous for the diagnosis, whether systemically modified or not. Refractory periodontitis, prepubertal periodontitis, and periodontitis associated with systemic disease are not recommended as useful descriptors of population samples without discrete identification of underlying systemic abnormality for all included subjects. Definition of population sample by a bacteriological or a host response feature is not recommended unless the trial is specifically aimed at that feature and the sample is homogeneous for it. All trials of efficacy should include physical or radiographic measurement of attachment level or bone height as a critical outcome variable. Results from trial in one form of periodontitis should not be applied directly to other forms.


Conduct of multicenter trials to test agents for treatment of periodontitis. The conduct of multicenter trials to test agents for treatment of periodontitis is conceptually similar to that of single center trials, but the requirement that centers ultimately be combined into a single result places much more stringent requirements on uniformity and control. Multicenter trials should be considered only if numbers of subjects or the ability to generalize to other treatment settings becomes a driving theme. General guidelines for the conduct of studies of this sort are suggested and problems that occur are discussed. Despite formidable difficulties that may occur, multicenter studies provide a convincing demonstration of an agent's therapeutic efficacy for the treatment of periodontitis.


Field testing as clinical trial methodology in periodontics. In the early 1950s the randomized control trial (RCT) was introduced and became widely accepted as the definitive proof of efficacy of a specific medical treatment. In fact, the acceptance and application of this methodology were instrumental in converting medicine from an unpredictable art to a science. At present no other methodologies exist that allow the evaluation of therapeutic efficacy with confidence comparable to that achieved with randomized controlled trials. In recent years researchers have applied new experimental designs and data analysis techniques to clinical trials conducted in a field trial environment to facilitate the understanding of proper use of new therapeutic agents and procedures. Since many of the new methodologies are still evolving or have only recently been introduced, this review considers some of the major trends and developments, as well as experiences of the authors, in field trial methodology. This manuscript addresses the following questions: 1) are there current clinical trial needs that are not met by RCT? 2) If so, what considerations are necessary for new approaches to have scientific usefulness? and 3) What are the strengths and weaknesses of the field trial's setting relative to an institutional environment?


Measurement of attachment level in clinical trials: probing methods. Measurement of clinical attachment level has become a standard for judging clinical response in periodontal therapy. A number of probing methods and instruments have been developed in an attempt to address limitations in obtaining this measurement. First generation instruments include conventional periodontal probes; second generation probes utilize controlled forces; and third generation probes incorporate automated measurement, controlled forces, and computerized data capture. Various types of stents have been used and repeated measurement techniques have been proposed to reduce examiner error. Controlled force probes appear to have their greatest advantage in increasing inter-examiner repeatability. The use of measurement stents increases inter- and intra-examiner reliability. However, use of such stents may be limited to small sample studies of limited duration. Third generation instruments offer advantages in terms of automated measurement and data capture, increased resolution, and a more continuous measurement scale, but do not necessarily result in increased intra- or inter-examiner reliability. Examiner training and calibration are essential for any measurement instrument. Decisions for or against use of a particular instrument must be made on the basis of the needs of each clinical trial. At the present time, no commercially available instrument resolves all of the inherent limitations of clinical measurement of attachment.


Radiographic methods in the evaluation of periodontal therapy. Radiographic methods are commonly utilized to assess periodontal bone changes in clinical trials. Radiographs are unique as a data source in that they provide a permanent visual record of the bone support and allow for linear, area, and volume measurements of periodontal lesions. The major limiting factors in the use of radiographic outcomes measures are geometric distortion and radiographic processing errors. The use of standardized methods and computer algorithms has reduced the effect of these errors on data from radiographs. Radiographic analysis in clinical trials is largely done in digital format by indirect digitizing film with a video camera. Direct digital detectors are now available for intra-oral use. The use of a direct detector eliminates the need for film processing and allows for dose reduction. The detectors currently available are limited in size and resolution. Subtraction radiography allows for area and volume estimates of bone gain or loss. The subtraction methods are highly precise, but are also technically exacting and labor intensive. The clinical trial hypothesis, size, length, and accuracy required should be the final consideration in choosing a radiographic method.


Methods for evaluation of regenerative procedures. A multitude of new procedures and materials are being tested for their ability to promote periodontal wound healing. This review provides an overview of the most well-accepted methods for evaluating periodontal wound healing procedures/materials. Topics discussed include appropriate patient selection, general principles in non-surgical and surgical therapy as they relate to wound healing trials, clinical soft and hard tissue measurements which are generally obtained, an overview of radiographic assessments, and a discussion of the methods and role of histological evaluations in clinical trials. The advantages and disadvantages of the various methodologies are presented. The review concludes with the potential endpoints which can be used in periodontal wound healing studies.


The host response in gingival crevicular fluid: potential applications in periodontitis clinical trials. Traditional clinical variables of periodontal pathology have only limited value as indicators for future disease progression in patients with adult periodontitis. Consequently, other aspects of the periodontal lesion are being examined for their diagnostic utility. Analysis of the host response in gingival crevicular fluid (GCF) is among the most intensely studied of these new diagnostic approaches. Specific indicators of the humoral immune response, cellular immune response, and acute inflammatory response have been identified in GCF. The relationship of indicators of the humoral immune response to active periodontal disease is equivocal. Specific indicators of the cellular immune response in GCF may ultimately prove to be important diagnostically, but the relationship of any specific marker to active periodontal disease has not been reported. In contrast, the acute inflammatory response in GCF has been extensively studied and a number of factors appear to be associated with an increased risk for future disease progression. Indicators of enhanced polymorphonuclear leukocyte activity, (lysosomal beta-glucuronidase, lysosomal collagenase), prostaglandin E2, and an indicator of acute tissue destruction (the cytoplasmic enzymes aspartate aminotransferase) have been associated with the occurrence of clinical attachment loss. An example of the application of a GCF marker in a periodontitis clinical trial is provided by describing the relationship of lysosomal beta-glucuronidase in GCF at baseline and 2 weeks following root planing and scaling to the occurrence of disease activity during the following 6 months. Persistently elevated levels of this enzyme were related to clinical attachment loss. The positive, negative, and total predictive values for beta-glucuronidase as an identifier of clinical attachment loss were 86%, 71%, and 76%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Design and analytic concepts for periodontal clinical trials. Aspects of the design and analysis of periodontal clinical trials are surveyed from a biostatistical perspective. Design issues discussed include protocol preparation, subject selection and its documentation, randomization, problems associated with the sample versus population paradigm in sampling of microbes and gingival fluid constituents, quality control, cross-over and split-mouth versus parallel-arm designs, blinding, and multicenter trials. Analytic discussion deals with the definition and choice of analytic unit, appropriate methods for the analysis of data from multiple sites within the same subject, the nature and application of randomization tests, interim analyses, subgroup analyses, and multiple comparison issues. Examples are provided to illustrate the feasibility of analyzing site-specific data while accounting for intra-subject correlation, which represents the increased similarity of sites chosen from the same mouth as compared to sites from different patients.


Inhibition of hepatic drug biotransformation by carrageenan-induced inflammation in the rat: effect of sex hormone alterations. Following in vivo treatment with carrageenan, sex-related differences in alteration of hepatic drug metabolism were found in the rat. In adult male rats, marked decreases were observed in hepatic 9000 x g supernatant cytochrome P-450 content and in the biotransformation of hexobarbital, aminopyrine, ethylmorphine, and meperidine. Hexobarbital hypnosis was significantly prolonged by carrageenan treatment in intact and testectomized animals as compared to their respective controls. Although carrageenan-treated intact animals slept 480% longer, carrageenan-treated testectomized rats slept only 60% longer than the respective control animals. However, testectomy or administration of 17 beta-estradiol to testectomized male rats did not inhibit the monooxygenase activities by carrageenan-treatment. Furthermore, administration of testosterone to ovariectomized female rats did not antagonize the inhibitory effects of the carrageenan-induced inflammation. The inhibitory effects produced by carrageenan-induced inflammation on the microsomal enzyme system were observed only in mature male rats and were not observed in mature female rats or in sexually immature rats of either sex. Thus, these results suggest that the inhibitory effects of carrageenan-induced inflammation on hepatic 9000 x g supernatant monooxygenases in the male rat are partially mediated through the toxic action of carrageenan-induced inflammation on androgen-dependent factors in this enzyme system.


Inhibition of hepatic mixed-function oxidase enzymes in mice by acute and chronic treatment with selenium. The effect of selenium administered acutely or chronically on the hepatic microsomal drug-metabolizing system has been investigated in mice. After 72 h following acute administration of selenium (7.5 mg/kg, i.p.), there was a significant inhibition of the activities of aminopyrine (AM) N-demethylase and ethylmorphine (EM) N-demethylase, and cytochrome P-450 levels but no change in the activities of aniline (AN) hydroxylase, 7-ethoxycoumarin (EC) O-deethylase, reduced nicotinamide adenine dinucleotide phosphate (NADPH)-cytochrome c reductase and reduced nicotinamide adenine dinucleotide (NADH)-ferricyanide reductase, and cytochrome b5 content. Chronic administration of selenium in the drinking water (1 or 2 ppm selenium) for 12 weeks, resulted in no alteration in any of the parameters measured. However, significant decreases in activities of AM N-demethylase and AN hydroxylase, and cytochrome P-450 levels were detected in animals given higher doses of selenium (4 or 8 ppm selenium). Following the in vitro additions of selenium to hepatic microsomes obtained from untreated mice, selenium inhibited the AM N-demethylase, AN hydroxylase and 7-EC O-deethylase in a concentration-dependent manner, but no alteration in NADPH-cytochrome c reductase and cytochrome P-450 levels was observed. These results indicate that selenium is a specific from inhibitor of hepatic monooxygenase.


[Conditions for analysis of cytochrome p-450 activity in human liver microsomes]. Multiples forms of CYT-P450 have been isolated from human liver microsomes. The distribution of CYT-P450 could be correlated with pathologic and influence the individual's response to therapeutic drugs and susceptibility to the toxic and cancerogenic effects of environmental pollutants. The aim of the this work is to determine the amounts of CYT-P450's in small samples from human liver such as biopsies and eventually to correlate them with pathology. For these reasons, tests providing informations about the distribution of CYT-P450 in individual subjects are very important. In this study we have reviewed the method for measuring the activity of CYT-P450 and the dosage of Benzopyrene hydroxylase. We modified some of these methods for the study of CYT-P450 from human liver microsomes.


[Autoimmune diseases: an aberrant behavior of our immune system?]. If autoimmunization seemed unthinkable at the beginning of this century, this phenomenon is now widely recognized. After a short recall of the immune system mechanism, we describe the recent concept allowing to explain the auto-antibody synthesis and their role in human pathology. The therapeutical strategies of autoimmune diseases are briefly evoked.


Species difference and tissue distribution of uridine diphosphate-glucuronyltransferase activities toward E6080, 1-naphthol and 4-hydroxybiphenyl. The apparent in vitro kinetic constants of uridine diphosphate-glucuronyltransferase (UDP-GT) activities towards E6080, 1-naphthol (1-N) and 4-hydroxybiphenyl (4-HB) were determined using microsomes, to assess the effect of inducing agents and evaluate species and tissue differences. In rats, the 3-methylcholanthrene and beta-naphthoflavone treatments increased the Vmax app/KM app values for E6080, 13- and 8-fold, and those for 1-N, 1.9- and 1.7-fold, respectively, but did not affect those for 4-HB. Phenobarbital was ineffective on the UDP-GT activity toward E6080. In rats and rhesus monkeys, the intestinal mucosa had higher specific UDP-GT activity toward E6080 than did the liver, and the rank order of the Vmax app/KM app value for E6080 in the intestinal mucosa was rhesus monkey > rat > guinea pig > beagle. In guinea pig, the Vmax app/KM app value for E6080 in the liver was 4 times higher than that in the intestinal mucosa. Both the Vmax app/KM app values for 1-N and 4-HB in the liver of all species studied were higher than those in the intestinal mucosa, and guinea pig liver showed the highest values, while, about the Vmax app for 1-N and 4-HB, beagle liver had the highest values. In the beagle intestinal mucosa, the Vmax app/KM app values for all 3 substrates studied were extremely low.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of cisapride on gastrointestinal motor activity and gastric emptying of disopyramide. The effects of cisapride on the gastrointestinal contractile activity and pharmacokinetics of disopyramide were determined in beagle dogs and patients with arrhythmia. In the animal experiments, the gastric motor index was significantly decreased by i.v. administration of disopyramide in a dose-dependent fashion. The peak decrease of the motor index was observed within 5 min after i.v. injection of disopyramide; the motor index then recovered gradually to the level present prior to drug administration. I.v. administration of cisapride (0.5 mg/kg) markedly increased gastrointestinal contractile activity following the decrease induced by disopyramide pretreatment (5 mg/kg, i.v.). In the clinical studies, the gastric emptying test was performed using the acetaminophen method. A significant correlation between plasma concentrations of disopyramide and gastric emptying time has been found (p < 0.001). The combination of disopyramide (100 mg t.i.d.) and cisapride (2.5 mg t.i.d.) significantly increased gastric emptying compared with that induced by disopyramide alone. The peak plasma concentration of disopyramide in association with cisapride oral administration was significantly higher, and the apparent absorption rate constant and lag time of disopyramide were about 2-fold higher and 2-fold shorter, respectively, than for disopyramide alone. Cisapride, acting as a cholinergic agonist, may counteract the anticholinergic effect of disopyramide on gastric motility. As a factor influencing drug absorption, gastric emptying is of importance, as it determines the rate of drug delivery to the small intestine. Therefore, the oral administration of disopyramide with cisapride may be useful for patients with delayed gastric emptying.


Chronopharmacological study of valproic acid in mice: comparison of oral and rectal administration. This study was performed to investigate the influence of the dosing route on chronopharmacological aspect of valproic acid (VPA) in mice, comparing the oral and rectal route. ICR male mice, housed under a standardized light-dark cycle (lights on from 0700 to 1900), were orally or rectally administered 400 mg/kg VPA each at the following scheduled time: 0900, 1300, 1700, 2100, 0100 and 0500. VPA concentrations in plasma and brain were determined by gas-liquid chromatography. There was a circadian rhythm in the electroshock seizure (ES) threshold 30 min after oral VPA administration, with the highest value at the midlight (1300) and the lowest at the middark (0100) (p < 0.01). A significant circadian rhythm was also found in plasma and brain VPA concentrations 30 min after oral administration (p < 0.01). This finding is related to the rhythm in the ES threshold. In contrast to oral administration, no circadian rhythm in the ES threshold, plasma and brain VPA concentrations was observed after rectal administration. These values after rectal dosing showed higher levels in comparison to those after oral dosing. Thus, the rectal route for VPA might have merit to eliminate the time-dependent changes in VPA pharmacologic action and kinetics. The timing of drug administration is an important factor that must be carefully controlled in drug pharmacokinetic and pharmacodynamic studies and must be considered in planning dosing routes.


Pharmacokinetics of zonisamide; saturable distribution into human and rat erythrocytes and into rat brain. The distribution of zonisamide, a new antiepileptic drug, in erythrocytes and in brain was studied to clarify the factors influencing its distribution in epileptic patients. In both humans and rats, zonisamide was concentrated significantly in erythrocytes in a saturable manner. When the effective concentration of zonisamide in serum was compared with that in blood in nine refractory epileptic patients taking zonisamide chronically, the variation in effective serum concentration was significantly larger than that in blood concentration. In rats, the distribution in the brain also showed saturability. These results suggest that differences in saturable binding to various tissues may contribute to the wide variation that occurs in the effective serum concentration of zonisamide in epileptic patients and that monitoring of the blood concentration of zonisamide may provide useful information for treatment with this drug.


Tissue distribution of intraperitoneally administered (1-->3)-beta-D-glucan (SSG), a highly branched antitumor glucan, in mice. Distribution of metabolically 3H-labeled (1-->3)-beta-D-glucan (3H-SSG) obtained from the culture filtrate of Sclerotinia sclerotiorum IFO 9395, in various tissues after intraperitoneal administration into ICR mice (250 micrograms/mouse) was examined. 3H-SSG was mainly detected in liver, spleen, and blood, and a negligible amount was excreted into the feces and excrement within 2 d. The significant amount of 3H-SSG remained in liver and spleen after 1 month. On the other hand, SSG was not incorporated effectively in vivo and in vitro by peritoneal exudate macrophages (0.5 microgram/1 x 10(6) M phi) Similarly in vivo, the majority of 3H-SSG distributed in spleen and liver were recovered from the non-cellular fraction and not from splenic macrophage and Kupffer cell fractions. These results suggested that (1-->3)-beta-D-glucans would not be easily incorporated by the host cells to degrade and exclude from the body even after the onset of the biological response modifier activity.


Evidence of direct conversion of testosterone sulfate to estradiol 17-sulfate by human placental microsomes. Ring A aromatization of testosterone sulfate (TS), using human term placental microsomes as a enzyme source, was examined under a reduced nicotinamide adenine dinucleotide phosphate (NADPH)-generating system. The aromatization to estradiol 17-sulfate (ES) without removal of the C-17 conjugate group was confirmed. Judging from the results of various experiments, including an inhibition experiment, the conversion was considered to occur by placental aromatase. Apparent Km and Vmax values for this aromatization were determined to be 37.4 microM and 330 pmol/mg protein/min respectively. The physiological role of direct conversion of TS by placenta to ES is discussed in connection with pregnancy.


Direct modulation of secretin binding sites by gastrin in the rat stomach. The effect of gastrin on secretin binding sites in the stomach was studied using the plasma membranes from rat gastric mucosa and vascularly perfused rat stomach. Tetragastrin transiently increased secretin binding to the mucosal plasma membranes. In the perfused stomach secretin binding was also modulated by the inclusion of tetragastrin or human [Leu15] gastrin I in the perfusate. However, histamine did not show such a modulatory effect. Tetragastrin had an insignificant effect on secretin binding sites in rat pancreas. These results suggest that the direct modulation of secretin binding by gastrin to its receptors may be involved in the inhibitory action of secretin on acid secretion induced by gastrin.


Development of physical dependence on morphine not accompanied with tolerance formation. The development of tolerance induced by daily repeated administration of 10 mg/kg of s.c. morphine was suppressed by combined injection of adrenergic blockers, 10 mg/kg i.p. phentolamine or propranolol, in mice. However, this treatment could not prevent the development of physical dependence as evidenced by the naloxone precipitated abstinence signs. Thus, we provide a new evidence that morphine analgesia, tolerance and physical dependence is dissociable from each other.


Homology of an acrosome-reacted sperm-specific antigen to CD46. MH61 is an acrosome-reacted sperm-specific monoclonal antibody. The antibody-coated beads effectively analyze the acrosomal status of human sperm. However, the nature of the antigen was not known. The antigen was purified by immunoprecipitation and SDS-PAGE followed by blotting on PVDF paper. The N-terminal sequence of the antigen was analyzed by an automated protein sequencer. An exactly matching sequence was found in CD46, that is also known as a membrane cofactor protein.


Family functioning when children have cystic fibrosis. The purpose of this study was to compare family functioning between 32 mothers with a child who had been diagnosed with cystic fibrosis (Group 1) and 32 mothers with a child who did not have cystic fibrosis (Group 2). The children's ages ranged from 4 months to 3 years. The mothers' perceptions of their family's functioning was measured by the Feetham Family Functioning Survey. There were no significant differences found between the groups on the total score of the Feetham Family Functioning Survey. Both groups of mothers reported that they were quite satisfied with their family functioning.


Nursing role supplementation for adolescent parents: prescriptive nursing practice. Role supplementation interventions were developed to support role taking in parenting and health decision-making for adolescent parents. The prescriptive nursing interventions provided during prenatal, well child, and home visits during the first 2 years of the child's life are based on role supplementation theory. Strategies derived from role theory are integrated with assessment and intervention activities. Adolescents receive role supplementation from a primary nurse who continues to see them and their infants over the 2-year period. The role supplementation intervention is currently being tested in a longitudinal research project.


Observation: a valuable data collection strategy for research with children. This study introduces the basic concepts of observational research by defining the terms and discussing some of the critical issues specific to observational research. Observational methods signify a systematic approach to quantifying behavior. A discussion of the major concerns in observational research is presented. These concerns include defining the coding categories; identifying the unit of measurement, the method of recording the observation, and deciding on a sampling strategy; training of observers; and interrater and intrarater reliability.


Sampling rare pediatric populations.

Sampling rare pediatric populations. The sampling of general populations of individuals or households for survey research is well described. However, much of the focus of pediatric nursing research is on special or rare populations. This article presents techniques that may be used to locate rare populations for pediatric nursing research. Additionally, development and analysis of one technique used to locate a sample frame for a survey of time use of chronically ill school-age children is discussed.


The creation, validation, and continuing development of the Oucher: a measure of pain intensity in children. This is the first article written by this group of authors/researchers who are collaborating on the development of the Oucher, an assessment tool to assist 3- to 12-year-olds describe the intensity of pain. The background and conceptual framework for its development, the research supporting the validation of the original Oucher, the research to create new ethnic versions, and the basic instructions for clinical use are described. In addition, several issues regarding the continued development and use of the Oucher are identified, including those relative to poster size and the gender and ethnicity of the photographed child. This article illustrates the precision and care needed to create clinically useful tools for obtaining information directly from young children.


The tendency for temperament to be "temperamental": conceptual and methodological considerations. Temperament seems to be a concept that stimulates a great deal of enthusiasm among nurses and professionals who "discover" it. Yet "knowing" that you have captured something of value does not necessarily imply full knowledge and understanding of the object. On the contrary temperament enthusiasts guarantee no consensus of opinion about what it is or how to best measure it. And so the conceptual and methodological issues for debate and discussion continue spilling over into both its research and clinical use. Sorting out the issues and the implications for use when there has been so much discussion and debate can be difficult. This article attempts to identify the main issues surrounding temperament, where we stand on some of the issues, and the practical implications for nurses who are interested in incorporating the measurement of temperament into clinical practice and research.


Pharmacoimmunodynamics of methylprednisolone: trafficking of helper T lymphocytes. A two-compartment closed model was used to characterize the cell trafficking behavior of helper T cells in response to various single doses of methylprednisolone. Steroids are assumed to inhibit the circadian-determined cell return from extravascular sites to blood in a classic inhibitory pattern reflected by an IC50. The rate of cell efflux from tissues is modeled with a cosine function having a period of 24 hr and a maximum at about 1 AM. Nonlinear least-squares regression employing differential equations was used to analyze helper T-cell data from three human studies from our laboratory. The IC50 value of methylprednisolone of 12-19 ng/ml approximates receptor KD values. Simulations were performed to demonstrate the log-linear role of steroid dose or AUC on the integral of effect of helper T cells over a wide range of methylprednisolone doses. This pharmacodynamic model allows flexibility for characterizing any type of steroid dosing regimen and is relevant in describing complex response data for corticosteroid immunosuppressive effects in man.


Biopharmaceutical evaluation of ibufenac, ibuprofen, and their hydroxyethoxy analogs in the rabbit eye. Two new structural analogs, 2-(4-hydroxyethoxyphenyl)acetic acid [R3] and 2-(4-hydroxyethoxyphenyl)propionic acid [R4], along with their parent compounds, ibufenac and ibuprofen, were evaluated for their biopharmaceutical properties. The analogs represented substitution of the lipophilic isobutyl side chains of ibufenac and ibuprofen with hydrophilic hydroxyethoxy side chains. Anti-inflammatory activity was evaluated by administering drugs topically to inhibit inflammation induced by using either clove oil or arachidonic acid. The rank order of activity was ibufenac approximately equal to ibuprofen > R3 approximately equal to R4. The new compounds, R3 and R4, were highly water soluble (> 60-fold) and partitioned less (< 1/1500-fold) into the lipid phase when compared to ibufenac and ibuprofen. R3 and R4 each had apparent corneal permeability coefficients of 6 x 10(-6) cm/sec, whereas ibufenac and ibuprofen yielded values of about 22 x 10(-6) cm/sec. In an ocular pharmacokinetic study in the rabbit eye, constant concentrations of each compound were maintained on the cornea in a cylinder or well fixed to the cornea, resulting in a constant input rate. This method circumvented parallel loss routes at the absorption site including nasolacrimal drainage. From area calculations the dispositions of the compounds within the eye were described by mean residence times, steady state volumes of distributions, and clearance rates. R3 and R4 were more slowly absorbed, retained within eye tissues longer, and were cleared more slowly from the eye than ibufenac and ibuprofen. The aqueous humor concentration-time profiles were also computer-fitted to equations representing classical pharmacokinetic models. For ibufenac and ibuprofen, the entire cornea was assumed to be the net barrier for entry into the anterior chamber. Whereas, for R3 and R4, the corneal epithelium and endothelium were presumed to be the diffusional barriers into and out of the stroma, the latter treated as a compartment. Aqueous humor concentrations of each drug fit the models reasonable well and agreed with conclusions made from the use of area calculations. The drop volume method was used to measure the surface tension of each compound. Both ibufenac and ibuprofen were considerably more surface active than R3 or R4. The greater surface tension measured for ibufenac and ibuprofen correlated to the subjective observations of ocular discomfort for these drugs.


Neural networks in pharmacodynamic modeling. Is current modeling practice of complex kinetic systems at a dead end? Neural networks (NN) are computational systems implemented in software or hardware that attempt to simulate the neurological processing abilities of biological systems, in particular the brain. Computational NN are classified as parallel distributed processing systems that for many tasks are recognized to have superior processing capability to the classical sequential Von Neuman computer model. NN are recognized mainly in terms of their adaptive learning and self-organization features and their nonlinear processing capability and are considered most suitable to deal with complex multivariate systems that are poorly understood and difficult to model by classical inductive, logically structured modeling techniques. A NN is applied to demonstrate one of the potentially many applications of NN for modeling complex kinetic systems. The NN was used to predict the effect of alfentanil on the heart rate resulting from a complex infusion scheme applied to six rabbits. Drug input-drug effect data resulting from a repeated, triple infusion rate scheme lasting from 30 to 180 min was used to train the NN to recognize and emulate the input-effect behavior of the system. With the NN memory fixed from the 30- to 180-min learning phase the NN was then tested for its ability to predict the effect resulting from a multiple infusion rate scheme applied in the subsequent 180 to 300 min of the experiment. The NN's ability to emulate the system (30-180 min) was excellent and its predictive extrapolation capability (180-300 min) was very good (mean relative prediction accuracy of 78%). The NN was best in predicting the higher intensity effect and was able to identify and predict an overshoot phenomenon likely caused by a withdrawal effect from acute tolerance. Current modeling philosophy and practice is discussed on the basis of the alternative offered by NN in the modeling of complex kinetic systems. In modeling such systems it is questioned whether traditional modeling practice that insists on structure relevance and conceptually pleasing structures has any practical advantages over the empirical NN approach that largely ignores structure relevance but concentrates on the emulation of the behavior of the kinetic system. The traditional searching for appropriate models of complex kinetic systems is a painstakingly slow process. In contrast, the search for empirical models using NN will continue to improve, limited only by technological advances supporting the very promising NN developments.


Passive tube and suction drainage after elective cholecystectomy--a comparison using ultrasonography. Daily ultrasonography of the gallbladder bed was performed in patients with suction or passive tube drains after elective cholecystectomy. A total of 19 patients was randomized to suction drainage and 17 to passive tube drainage. A policy of early drain removal was followed. No significant difference was found between the volume drained and the size of collection detected in either group. Significant bile leaks were detected and were adequately drained by suction and passive tube drains. There were no complications from drains. In view of these findings, we advocate short-term drainage of the gallbladder bed after both open and laparoscopic cholecystectomy using the drain of the surgeon's choice.


Ovulation induction with human menopausal gonadotropin versus follicle-stimulating hormone after pituitary suppression by gonadotropin releasing hormone agonist in polycystic ovary disease. A cross-over study. Ten patients with polycystic ovary disease (PCOD) had ovulation induction after pituitary suppression by gonadotropin releasing hormone agonist (GnRHa) with GnRHa plus pure follicle-stimulating hormone (FSH) or plus human menopausal gonadotropin (hMG). Duration of the stimulation period and gonadotropin doses were superimposable. A multifollicular response was observed in both treatments. Bioassay and radioimmunoassay of luteinizing hormone, androstanedione and testosterone plasma levels were higher in hMG cycles compared to FSH-treated cycles. No differences was found in FSH and estradiol (E2) plasma concentrations, whereas in hMG-treated cycles the E2/number of follicles and E2/ovarian volume ratios were greater than in the FSH-treated cycles. Clinical results in terms of percentages of ovulation and pregnancies were the same in the two protocols. We conclude that the presence of luteinizing hormone in induction of ovulation in patients with PCOD does not seem to influence follicular recruitment and development, but it may have a role in the enhancement of steroid production.


Maternal hepatitis B screening at a private hospital. A prospective study was performed to determine whether the Centers for Disease Control risk factors are reliable predictors of the hepatitis B surface antigen (HBsAg) carrier state in the obstetric population at a large private hospital in Cincinnati. During the 12-month study period, 5,877 patients delivered at the hospital. The patients were screened for HBsAg either prenatally or on presentation in labor. Questionnaires were administered after arrival at the hospital to assess for historical risk factors. An overall 0.0925% incidence of HBsAg seropositivity was discovered. All patients who were HBsAg positive had identifiable risk factors.


Recurrence rate after fluid aspiration from sonographically benign-appearing ovarian cysts. The outcome after fluid aspiration from 41 sonographically benign-appearing ovarian cysts was assessed. The considerable probability of recurrence was significantly higher after ultrasound-guided aspiration than following aspiration via laparoscopy (54% versus 30%, respectively, at 36 months). Although all the recurring cysts were benign, the concern with malignancy and the high recurrence rate seem to indicate that fluid aspiration from sonographically benign-appearing cysts by either method is not the management of choice.


Zygosity and umbilical cord length.

Zygosity and umbilical cord length. Differences in cord length between weight-concordant pairs were studied in 43 monochorionic, 59 same-sex dichorionic and 42 different-sex dichorionic sets of twins and comparisons were made between groups. Of obvious and curious note was the narrow range of cord length differences between twins in monochorionic pairs (range, 0-5 cm; mean, 2.1 +/- 1.6 cm). Same-sex and different-sex dichorionic groups had significantly wider ranges of cord length differences than the monochorionic group (range, 0-34 cm; means, 7.7 +/- 6.4 and 11.0 +/- 6.1 cm, respectively). In the same-sex dichorionic group no pair whose cord lengths differ by > 5 cm has yet been found to be identical. Discordant monochorionic twins had greater differences in cord length. All other factors being equal, the remarkably similar cord lengths of concordant identical twins strongly supports the hypothesis that a genetic component influences cord length.


Decreasing risk of pregnancy loss following chorionic villus sampling. Elimination of transabdominal chorionic villus sampling during the ninth week of pregnancy. Chorionic villus sampling (CVS) is a method of obtaining fetal cells in the first trimester of pregnancy for genetic analysis. The transcervical (TC) approach was the first technique to be widely used. In the National Institute of Child Health and Human Development collaborative study the absolute loss rate following CVS (the total number of spontaneous abortions and neonatal deaths following CVS) was 4%. More recently the transabdominal (TA) approach has been introduced. This study compares the loss rates for the two approaches at various gestational ages for three 6-month periods following the addition of the TA approach with each other and with the loss rates prior to the introduction of TA CVS. We found that the percentage of pregnancy losses following TA CVS during the ninth week of gestation (63-69 days) was consistently higher than for TC CVS performed at the same gestational age. The loss rate for TC CVS has steadily decreased since the introduction of TA CVS after remaining the same for the two years prior to the introduction of the TA approach. After minimizing the number of TA CVS performed during the ninth week of gestation, the overall loss rate during the most recent 6-month period has been reduced to 0.94%. We conclude that the lowest loss rate following CVS can be obtained if both the TA and TC methods are available, and that the number of TA procedures performed during the ninth week of gestation is minimized.


Breast evaluation and diagnosis by obstetrician-gynecologists. Survey of practice patterns. Surveys of the obstetrician-gynecologists attending the breast disease postgraduate courses at the 1988 and 1990 American College of Obstetricians and Gynecologists annual clinical meetings report a high level of practice involvement in breast disease evaluation and cancer screening for their patients. A consistently high percentage stated they performed regular clinical breast examinations, documented the examinations with a diagram in the medical record, gave breast self-examination instruction, advised screening mammography following the American College of Obstetricians and Gynecologists guidelines, utilized a patient tracking system for follow-up and referred patients with undiagnosed dominant breast masses. Most stated that they did breast cyst aspiration; those who did not mostly referred their patients for cyst aspiration. The accurate and cost- and time-effective office technique of fine needle aspiration of palpable dominant solid breast mass continues to be underutilized even though the procedure has been proven effective and accurate in the cytologic diagnosis of benign and malignant breast neoplasms. Instruction in the technique of fine needle aspiration of palpable breast cysts and solid masses is available in many clinics, workshops and postgraduate courses.


Survey of colposcopy practices by obstetrician/gynecologists. A statewide survey to characterize the colposcopy patterns of practicing obstetrician/gynecologists was undertaken. There was a 66.1% response rate, with 98.2% of respondents performing colposcopy. Of those performing colposcopy, nearly all perform biopsies, cryosurgery and conizations; 73.4% perform laser vaporization, 66.7% perform laser cone biopsy and 11% perform laser cone biopsies in their offices. The mean number of colposcopies performed by respondents in a six-month period was 55. Twenty percent performed less than one examination per week and an additional 60% performed two to three examinations per week. Further studies to assess the diagnostic accuracy of those performing greater and lesser numbers of examinations are needed.


Cumulative pregnancy rates in patients with apparently normal fertility and fertility-focused intercourse. Fifty consecutive clients achieved pregnancy using a standardization modification of the Billings ovulation method (the Creighton Model Natural Family Planning System). Of 50 clients followed, 38 (76%) achieved pregnancy in the first cycle of fertility-focused intercourse, 45 of 50 (90%) achieved pregnancy by the third cycle and 49 of 50 achieved pregnancy by the sixth cycle (98%).


Intrapartum amniotic fluid index and neonatal acidosis. A pilot study to determine the correlation. The purpose of a prospective study was to determine if there was a significant association between an intrapartum amniotic fluid index (AFI) < or = 5.0 cm and neonatal acidosis (umbilical arterial pH < 7.20 or metabolic acidosis [umbilical arterial pH < 7.20 and base deficit > 10 mEq/L]). In early labor 101 gravidas at > or = 37 gestational weeks underwent a four-quadrant amniotic fluid assessment; at delivery, umbilical arterial acid-base levels were determined. Among women with AFI < or = 5.0 cm as compared to those with AFI > 5.0 cm, the fetuses were more likely to have neonatal acidosis (31.2% versus 17.6%, respectively) and metabolic acidosis (25.0% versus 10.5%, respectively), although the differences were not statistically significant.


Effects of sex steroids on cell differentiation and interleukin-1 beta production in the human promyelocytic leukemia cell line HL-60. The effects of 17 beta-estradiol (E2), progesterone (P) and testosterone (Te) on cell differentiation in the HL-60 promyelocytic leukemia cells after treatment with 1 alpha, 25-dihydroxyvitamin D3 [1,25-(OH)2D3] and those on interleukin-1 beta (IL-1 beta) production by HL-60 cells in response to lipopolysaccharide (LPS) were investigated. Neither E2 (10(-10) to 10(-7) M), P (10(-9) to 10(-6) M) nor Te (10(-10) to 10(-7) M) affected monocytic differentiation as assessed by reactivity with OKM14 monoclonal antibody and alpha-naphthyl acetate esterase activity. Pretreatment of HL-60 cells with 1,25-(OH)2D3 enhanced their ability to produce IL-1 beta in response to subsequent exposure to LPS, although 1,25-(OH)2D3 by itself did not induce IL-1 beta production by HL-60 cells. This priming effect of 1,25-(OH)2D3 was augmented by the addition of E2 and Te at physiologic concentrations, but not by that of P. E2, P and Te at physiologic concentrations enhanced IL-1 beta production by HL-60 cells that were pretreated with 1,25(OH)2D3 and stimulated by LPS. The increasing rate of IL-1 beta production by the addition of E2 and Te was higher when added with LPS than when added with 1,25-(OH)2D3. These findings suggest that enhancing effects of sex steroids in IL-1 beta production by monocyte/macrophage lineage cells.


Vaginal birth after cesarean section at the University of Texas. Vaginal delivery was successful in 76% of the 242 women who underwent a trial of labor after cesarean section in a prior pregnancy. Separation of the uterine scar occurred in four women (1.7%). Women whose prior cesarean section was for breech presentation had the highest rate of successful vaginal delivery (86%). The vaginal delivery rates were similar in women who delivered infants with birth weights > or = 4,000 g (73%) and < 4,000 g (76%). The use of epidural anesthesia and oxytocin may enhance the success of vaginal delivery in women undergoing a trial of labor following an earlier cesarean section.


Iatrogenic omental percutaneous shunt and resolution of obstructive fetal uropathy. A case report. A double pigtail stent was placed to decompress an obstructed fetal kidney. The stent was dislodged, causing an iatrogenic marsupialization between the renal pelvis omentum and skin. The omentum acted as a drain, decompressing the kidney.


Iniencephaly. A case report.

Iniencephaly. A case report. Prenatal ultrasound diagnosed iniencephaly apertus at 21 weeks' gestation. In this rare central nervous system (CNS) malformation the brain and neck show the main pathologies. Retroflexion of the head with exaggerated cervicothoracic lordosis is always present, and CNS malformations in the form of anencephaly, spina bifida and encephalocele are often present. The ultrasonic diagnosis should be based on the finding of extreme dorsiflexion of the head accompanied by an abnormally short and deformed spine.


Advanced squamous cell carcinoma of the vulva after treatment for verrucous carcinoma. A case report. Verrucous carcinoma of the vulva, a rare variant of squamous cell carcinoma, is a locally destructive, nonmetastasizing tumor. While controversy surrounds the choice of treatment for this lesion, which frequently recurs, it is generally accepted that a recurrence is almost always local, with histologic characteristics and biologic behavior the same as those of the primary tumor. A case of squamous cell carcinoma of the vulva with involvement of the regional lymph nodes after treatment of verrucous carcinoma is reported to increase awareness of the potential biologic behavior of this uncommon tumor.


Placenta increta complicating a first-trimester abortion. A case report. Placenta increta complicating pregnancy in the first trimester is rare. A patient with risk factors for placenta increta required a hysterectomy to control a hemorrhage after a first-trimester abortion. Pathologic study confirmed the preoperative diagnosis of placenta increta.


Mortality following unilateral twin interstitial ectopic pregnancy. A case report. Twin ectopic pregnancy is an uncommon event, usually occurring as simultaneous intrauterine and tubal gestations. Interstitial implantation of an ectopic pregnancy is also a rare event, associated with a high mortality rate. Twin interstitial pregnancy has been previously reported only three times in the English literature. We report a recent case of unilateral twin interstitial ectopic pregnancy that resulted in maternal death and review the literature with regard to both ectopic pregnancy and factors associated with mortality from interstitial implantation.


2-(aminoalkyl)-5-nitropyrazolo[3,4,5-kl]acridines, a new class of anticancer agents. 2-(Aminoalkyl)-5-nitropyrazolo[3,4,5-kl]acridines were prepared from substituted anilines via the 1-chloro-4-nitroacridones followed by condensation with [(alkylamino)alkyl]hydrazines. Impressive activity was demonstrated for the 9-hydroxy, 9-alkoxy, and 9-acyloxy analogs in vitro on a L1210 leukemia line and in vivo against the P388 leukemia. Advanced studies led to the selection of 3bbb for clinical trial.


Synthesis and physicochemical properties of sulfamate derivatives as topical antiglaucoma agents. Several imidazolylphenyl sulfamate and (imidazolylphenoxy)alkyl sulfamate derivatives were synthesized and evaluated as topically active carbonic anhydrase inhibitors. Water solubility, pKa, carbonic anhydrase inhibition, and partition coefficient for the compounds were measured. Sulfamic acid 2-[4-(1H-imidazol-1-yl)phenoxy]ethyl ester monohydrochloride (16) has the best combination of properties and showed excellent topical activity in lowering the intraocular pressure in New Zealand white rabbits.


Inhibition of human leukocyte elastase by N-substituted peptides containing alpha,alpha-difluorostatone residues at P1. A series of tripeptides which contain alpha,alpha-difluorostatone residues at P1-P1' and span the S3-S1' subsites have been shown to be potent inhibitors of human leukocyte elastase (HLE). The tripeptides described contain the nonproteinogenic achiral residue N-(2,3-dihydro-1H-inden-2-yl)glycine at the P2-position. This redidue has previously been shown in the case of HLE to be a good bioisosteric replacement for L-proline. Of the peptides prepared, those which contain the alpha,alpha-difluoromethylene keton derivative of L-valine (difluorostatone) are the preferred residue at the P1-primary specificity position. Substitution at P1 by the corresponding alpha,alpha-difluoromethylene ketones of L-leucine and L-phenylalanine gives inactive compounds. Of the tripeptides described the most potent in vitro compound is ethyl N-[N-CBZ-L-valyl-N-(2,3-dihydro-1H-inden-2-yl)glycyl]- 4(S)-amino-2,2-difluoro-3-oxo-5-methylhexanoate (17B) (IC50 = 0.635 microM). It is presumed that the inhibitor 17b interacts with the S3-S1' binding regions of HLE. Additionally extended binding inhibitors were prepared which interact with the S3-S3' binding subsites of HLE. In order to effect interaction with the S1'-S3' subsites of HLE, the leaving group side of cleaved peptides, spacers based upon Gly-Gly, and those linked via the N epsilon of L-lysine were utilized. One of the most potent extended compounds (P3-P3') in vitro is methyl N6-[4(S)-[[N-[N-CBZ-L-valyl-N- (2,3-dihydro-1H-inden-2-yl)glycyl]amino]-2,2-difluoro-3-oxo-5- methylhexanoyl]-2(S)-(acetylamino)-6-aminohexanoate (24b) (IC50 = 0.057 microM). The described in vitro active inhibitors were also evaluated in hamsters in an elastase-induced pulmonary hemorrhage (EPH) model. In this model, intratracheal (it.) administration of 22c, 5 min prior to HLE challenge (10 micrograms, it.) effectively inhibited hemorrhage (94.6%) in a dose-dependent manner. The described alpha,alpha-difluoromethylene ketone inhibitors are assumed to act as transition-state analogs. The inhibition process presumably acts via hemiketal formation with the active site Ser195 of HLE, and is facilitated by the strongly electron withdrawing effect of the alpha,alpha-difluoromethylene functionality.


Antimitotic agents: ring analogues and derivatives of ethyl [(S)-5-amino-1,2-dihydro-2-methyl-3-phenylpyrido[3,4-b]pyrazin-7- yl]carbamate. The synthesis of ring analogues and derivatives of the S isomer of ethyl [5-amino-1,2-dihydro-2-methyl-3-phenylpyrido[3,4-b]pyrazin-7 - yl[carbamate, (S)-1, a potent antimitotic agent with anticancer activity, was directed toward the determination of the contribution of several structural features of this compound to biological activity. Replacement of the 5-amino with a 5(6H)-oxo group and either transposing the 6-ring nitrogen to or incorporation of a ring nitrogen at the 8-position caused a significant decrease in in vitro activity and destroyed in vivo activity. Although in vivo cytotoxicity was reduced, in vitro activity at higher doses relative to (S)-1 was retained by replacement of the 5-amino group with hydrogen and by expansion of the 1,2-dihydropyrazine to give a dihydro-1,4-diazepine ring.


Hypoxia-selective antitumor agents. 6. 4-(Alkylamino)nitroquinolines: a new class of hypoxia-selective cytotoxins. A series of isomeric 4-[[3-(dimethylamino)propyl]amino]nitroquinolines has been synthesized and evaluated as hypoxia-selective cytotoxins and as radiosensitizers of hypoxic cells. The compounds showed widely-differing hypersensitivity factors (ratios of cytotoxicity against wild-type and repair-deficient mammalian cells). Many compounds showed oxygen-sensitive bioreduction resulting in DNA alkylation, while others show oxygen-insensitive modes of action. Of the nitro isomers studied, the 5-nitro showed the greatest hypoxic selectivity. A series of ring-substituted analogues were then prepared, in an effort to lower its reduction potential of -286 mV. Structure-activity studies showed that the effects of substitution on reduction potential were complex, being mediated by electronic and steric effects on the nitro group, as well as by effects on quinoline pKa. Two compounds of lower reduction potential, the 3- and 8-methyl analogues, showed improved selectivity (47- and 60-fold in a clonogenic assay). These two compounds also showed the highest "in vitro therapeutic indices" of the series as hypoxic cell radiosensitizers. Despite these favorable in vitro properties, neither compound had activity against hypoxic cells in SCCVII tumors when administered at 60% of the MTD.


The conformation of 6-methoxyerythromycin A in water determined by proton NMR spectroscopy. The proton NMR spectrum of an aqueous solution of 6-methoxyerythromycin A (2) has been assigned and nuclear Overhauser effects have been obtained from a series of NOESY spectra. Carbon-13 antisymetric spin-lattice relaxation times have been measured for the methylene and methine carbons. The NMR data show that 2 is present exclusively in the keto form and has a conformation very similar to that reported for erythromycin A (1) in the solid and solution phases.


N-methylcarbamate derivatives of ellipticine and olivacine with cytotoxic activity against four human lung cancer cell lines. A series of analogues of the antitumor alkaloids ellipticine and olivacine were tested for cytotoxicity against four human lung cancer cell lines: H69, N417, H460, and H358. Adriamycin (doxorubicin), ellipticine, olivacine, and celiptinium were used as standards. Adriamycin was cytotoxic at 2 microM and celiptinium was inactive at the highest concentrations tested (IC50 > 48 microM). N-methylcarbamates of 9-methoxy-6H-pyrido[4,3-b]carbazole 1-,5-, and 11-methanols gave IC50 values ranging from 0.02 to 0.11 microM against N417, H460, and H358 and were only slightly less effective against H69.


New bronchodilators. 3. Imidazo[4,5-c][1,8]naphthyridin-4(5H)-ones. In order to develop new oral bronchodilators, a series of novel imidazol[4,5-c][1,8]naphthyridin-4(5H)-ones 5 were designed and synthesized. Some of these new heterocycles exhibited more potent bronchodilator activity in vitro and in vivo than theophylline. With respect to modification at the 5-position, both phenyl and n-butyl substitution produced potent activity. Though bulk tolerance at N-3 is observed with short and small lipophilic groups, any substitution at the other positions and transformations of the parent skeleton eliminated activity. Thus 5-phenyl-1H-imidazo[4,5-c][1,8]naphthyridin-4(5H)-one (23) (KF17625), which satisfied these conditions, was selected for further studies (antigen inhalation-induced bronchospasm model; minimum effective dose (MED) = 1 mg/kg, po; antigen-induced contraction of trachea (the Schultz-Dale reaction), IC50 = 2.2 microM). Compound 23 inhibited carbachol-, histamine-, or leukotriene D4-induced contraction and relaxed spontaneous tone in guinea pig isolated tracheal preparations with, 4- to 16-fold greater potency than aminophylline. Thus it appeared to relax directly the airway smooth muscle. 23 did not have any influence on adenosine binding at 10 microM, but inhibited canine tracheal phosphodiesterase (PDE) IV (IC50 = 12 microM) and concanavalin-A-induced histamine release from rat mast cells (44% inhibition at 10 microM). Although the detailed mechanisms of these compounds remain to be elucidated, this series of novel tricyclic heterocycles represents a new class of bronchodilator.


ATP-citrate lyase as a target for hypolipidemic intervention. Sulfoximine and 3-hydroxy-beta-lactam containing analogues of citric acid as potential tight-binding inhibitors. Citric acid analogues (+/-)-12a,b and (+/-)-17a,b, where one of the primary carboxylates has been replaced by a sulfoximinoyl and a 3-(3-hydroxy-beta-lactamyl) moiety, respectively, have been synthesized and evaluated as inhibitors of ATP-citrate lyase. The design of these inhibitors was based on methionine sulfoximine and tabtoxinine beta-lactam, potent, tight-binding inhibitors of glutamine synthetase. Both ATP-citrate lyase and glutamine synthetase employ phosphate-carboxylate anhydrides as a method for carboxylate activation during catalysis. Only one diastereomer, (+/-)-12a, displayed weak, reversible inhibition, while the remaining citrate analogues (+/-)-12b and (+/-)-17a,b were inactive against the lyase. No time-dependent inactivation of the enzyme was observed.


Alpha-halo [(phenylphosphinyl)methyl]phosphonates as specific inhibitors of Na(+)-gradient-dependent Na(+)-phosphate cotransport across renal brush border membrane. Certain phosphonocarboxylate analogues of phosphate are known to inhibit Na(+)-phosphate (Pi) cotransport in renal brush border membrane (BBM), but previously tested potential inhibitors incorporating structurally versatile aryl functionality were inactive. In this work, a series of novel alpha-halogenated [(phenylphosphinyl)methyl]phosphonates [PhpXYMP: X, Y = H, F (2); F, F (3); H, Cl (6); Cl, Cl (4); H, Br (7); Br, Br (5); and Cl, Br (8)] were prepared via synthesis of the corresponding triethyl esters, acid hydrolysis, and isolation as pyridine salts. The compounds were evaluated as inhibitors of Na(+)-gradient-dependent 32Pi uptake by rat renal cortex BBM vesicles (BBMV) in vitro. The PhpFMP racemate 2 had higher activity (-49% delta inhibition) than other members of the series (-22 to -39% delta inhibition). pKa values of 1.5-2.0, 2.7, and 7.1 were estimated for 2 using a 31P delta vs pH plot, indicating that in the activity assays it exists as both dianion and trianion, with the latter form predominant. PhpFMP had no significant inhibitory effect on Na(+)-gradient-dependent uptake of D-glucose or L-proline in the same BBMV, and did not inhibit BBM alkaline phosphatase. Kinetic analysis showed that PhpFMP acts as a strictly competitive inhibitor of Na(+)-Pi cotransport with Ki = 0.358 +/- 0.021 mM (n = 3). The racemate 2 was resolved as its (-)-quinine salt into enantiopure (+)-2 [Na+ salt, [alpha]25D = +6 degrees (aqueous MeOH)] and a Na+ salt of 2 enriched in (-)-2. The two compounds did not differ significantly as inhibitors of Na(+)-gradient dependent 32Pi uptake by rat renal cortex BBM vesicles (BBMV) in vitro. The results are discussed in terms of structural requirements for inhibition.


5-HT3 receptor antagonists. 1. New quinoline derivatives. A series of esters and amides of 1-alkyl-2-oxo-1,2-dihydroquinoline-4- carboxylic acid or 2-alkoxy-quinoline-4-carboxylic acid containing a basic azabicycloalkyl moiety has been synthesized and evaluated for affinity for the [3H]quipazine-labeled 5-HT3 receptors. Most of the esters exhibited 10-fold more potent activity than that of ondansetron (1; Ki = 7.6 nM). Lipophilic substituents at the 1- or 2-position of the quinoline ring enhanced affinity for the receptors. Compounds 21 and 37 showed the highest affinity (Ki = 0.32 and 0.31 nM, respectively) among them. On the other hand, most of the amides showed 100-fold lower affinity than that of the esters. Molecular modeling studies indicated that the carbonyl moiety in 19 (ester) or 31 (amide) was not coplanar to the plane of an aromatic ring (over 20 degrees deviation). Although some of the selected compounds exhibited potent activity in the Bezold-Jarisch (B-J) reflex test, good correlation was not observed between the affinity for the 5-HT3 receptors and the activity in the B-J reflex test (in vivo). From these data, it was suggested that our quinoline derivatives might interact with the 5-HT3 receptors in a different way from that of the reported 5-HT3 receptor antagonists presumably due to the presence of the heterogeneity of the 5-HT3 receptors between brain and heart.


Ketanserin analogues: structure-affinity relationships for 5-HT2 and 5-HT1C serotonin receptor binding. Ketanserin is the prototypic 5-HT2 serotonin antagonist; although it has been an important tool for the study of serotonin pharmacology, it has had relatively little impact on drug design because remarkably little is known about its structure-affinity relationships. Furthermore, ketanserin also binds at 5-HT1C receptors and even less is known about the influence of its structural features on 5-HT1C receptor affinity. The present study reveals that the fluoro and carbonyl groups of the 4-fluorobenzoyl portion of ketanserin make small contributions to 5-HT2 binding and that the intact benzoylpiperidine moiety is an important feature. Ring-opening of the piperidine ring reduces affinity. Although the quinazoline-2,4-dione moiety also contributes to binding, it appears to play a smaller role and can be structurally simplified with retention of 5-HT2 affinity. N-(4-Phenylbutyl)-4-(4-fluorobenzoyl)piperidine (39), for example, binds with nearly the same affinity (Ki = 5.3 nM) as ketanserin (Ki = 3.5 nM). All of the compounds examined bind at 5-HT1C sites with lower affinity than ketanserin, and some of the simplified analogues bind with nearly 10 times the 5-HT2 versus 5-HT1C selectivity of ketanserin; however, none displays > 120-fold selectivity. Several of the compounds, such as the amide 32 and the urea 33 represent examples of new structural classes of 5-HT2 ligands.


Modulation of aortic protein phosphorylation by benzo(a)pyrene: implications in PAH-induced atherogenesis. The toxicity of polycyclic aromatic hydrocarbons such as benzo(a)pyrene, 7,12-dimethylbenz(a)anthracene, and 3-methylcholanthrene has been associated with alterations in the proliferation of vascular smooth muscle cells and the development of lesions of mesenchymal origin. Because phosphorylation of endogenous substrates plays a central role in the regulation of smooth muscle cell growth, the present studies were conducted to evaluate the phosphorylation pattern of medial aortic protein upon repeated in vivo exposure of Japanese quail to benzo(a)pyrene (BaP). Medial aortic homogenates from quail treated for 10 weeks with 10 mg/kg benzo(a)pyrene or vehicle were processed for in vitro measurements of protein phosphorylation. In vitro phosphorylation of endogenous or exogenous proteins stimulated in vitro by phorbol myristate acetate/phosphatidylserine or cyclic AMP, known activators of protein kinase C and cyclic AMP-dependent protein kinase, respectively, was examined in the cytosolic and particulate fractions of homogenates from control and treated animals. Benzo(a)pyrene treatment significantly enhanced the basal phosphorylation of M(r) 113, 35, and 23 kDa proteins in the cytosolic fraction. Modest increases in the phosphorylation of M(r) 71, 52, and 38 kDa were also observed under basal conditions. No changes in the basal phosphorylation of particulate proteins were observed. Phosphorylation of endogenous protein substrates by protein kinase C in the cytosolic fraction was not altered by benzo(a)pyrene treatment. In contrast, inhibition of C-kinase-mediated phosphorylation of endogenous M(r) 272, 72, and 45 kDa proteins was observed in the particulate fraction of aortic homogenates from benzo(a)pyrene-treated quail relative to controls. Exogenous histone phosphorylation by PKC in the particulate, but not cytosolic fraction, was decreased by benzo(a)pyrene treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


Additive effects and potential inhibitory mechanism of some common aromatic pollutants on in vitro mitochondrial respiration. The in vitro toxicity of multiple hydrophobic compounds was the focus of this study. A mitochondrial respiratory assay, sensitive to perturbations caused by hydrophobic chemicals, was utilized to measure the effects of individual aromatic hydrocarbon pollutants and their mixtures on mitochondrial respiratory function. Benzene, naphthalene, acenaphthene, and 1-chloronaphthalene, common industrial solvents shown to interact additively in vivo, were evaluated using this in vitro assay system. Mitochondrial respiration was inhibited 50% (EC50) by 525 ppm (6.7 mM) benzene, 15 ppm (117 microM) naphthalene, 3.9 ppm (25.5 microM) acenaphthene, or 3.8 ppm (23.4 microM) 1-chloronaphthalene. NADH:O2 oxidoreductase (NADH-->O2), NADH:ubiquinone oxidoreductase, and ubiquinol:O2 oxidoreductase activities were inhibited by all four compounds, whereas succinate:O2 oxidoreductase, cytochrome c oxidase, and duroquinol:O2 oxidoreductase activities were not inhibited. Inhibition of mitochondrial respiration occurred at the level of ubiquinone (coenzyme Q10) for all four aromatic hydrocarbons. The ultraviolet absorbance spectrum of isolated Q10 was also altered by naphthalene, acenaphthene, or 1-chloronaphthalene, suggesting a specific interaction between that component of the respiratory chain and these aromatic hydrocarbons. Inhibition by a mixture of 2, 3, or 4 of the compounds tested was additive, reflecting a summation effect of each compound present in the mixture. This additive nature is consistent with previously reported effects of these compounds in vivo and with compounds having similar modes of action. The similar mode of action in vitro is a specific interaction with coenzyme Q10, not a generalized membrane perturbation as speculated to occur in vivo, and is the likely mechanism for the observed additive toxicity.


The flavin-containing monooxygenase in mouse lung: evidence for expression of multiple forms. The flavin-containing monooxygenase (FMO) was purified from mouse lung microsomes. On SDS-PAGE, the purified enzyme separated as two bands, a major band of 58,000 daltons and a minor band of 59,000 daltons. Antibodies to mouse liver FMO cross-reacted with both bands in the purified preparations, whereas antibodies to rabbit lung FMO cross-reacted only with the major band. In microsomal preparations the major band was recognized by both antibodies, but neither antibody detected the minor band in microsomes. A cDNA encoding the pig liver FMO hybridized with mRNA isolated from mouse liver, kidney, and lung, whereas cDNA encoding the rabbit lung FMO hybridized only with mouse lung and kidney mRNA. Thermal stability studies showed that the FMO preparation purified from mouse lung consisted of a heat-stable and a heat-labile component. The heat-labile component of lung FMO was inhibited competitively by imipramine, whereas the heat-stable component was insensitive to the presence of imipramine. Immunoprecipitation of purified mouse lung FMO with anti-rabbit lung FMO completely removed the protein band reactive to anti-rabbit lung FMO while leaving reactivity to anti-liver FMO. The catalytic and immunochemical differences seen between FMO from rabbit lung and mouse lung appear to result from the expression of at least two forms of FMO in the mouse lung, one similar to the rabbit pulmonary form and one similar to the major mouse liver form of FMO.


Regional distribution of neuropeptide-degrading enzyme activity in the rat brain: effects of subacute exposure to carbon disulfide. Carbon disulfide, a volatile solvent, is widely used in industry. It has been demonstrated that it causes several neuropsychological symptoms. However, the neurochemical basis of its neurotoxic effect is relatively unknown. In this paper we have measured the effect of subacute i.p. administration on neutral and basic aminopeptidase activities in discrete zones of the rat brain using lysine- and leucine-2-naphthylamides as substrates. Neutral aminopeptidase activity showed a significant decrease in the thalamus and cerebellum with marked (not significant) changes in the hypothalamus, hippocampus, medulla, and occipital cortex. There were no changes in basic aminopeptidase activity. It is suggested that aminopeptidase activity could play a role in carbon disulfide neurotoxic action in the aforementioned regions by generating changes in several neuropeptide levels.


Action of some retinol derivatives and their provitamins on microsome-catalyzed formation of benzo[a]pyrene-DNA adduct. Several vitamin A compounds have been tested for their ability to suppress formation of DNA adduct by the carcinogen benzo[a]pyrene (B[a]P) in an in vitro reaction catalyzed by rat liver microsomes. Retinol, retinal, 3-dehydroretinol and 3-hydroxyretinol were found to be effective inhibitors of adduct formation. Certain carotenoids that are precursors of these retinoids also displayed considerable inhibitory capacity. Carotenoids and the 3-substituted retinoids appeared to modulate the DNA adduct formation exclusively through their action on microsomal enzymes, since an effective inhibition in each case was observed on the formation of B[a]P-7,8-diol, a proximate carcinogenic metabolite of B[a]P. Unsubstituted retinoids, on the other hand, had marginal effect on enzymes but were found effective in accelerating inactivation of B[a]P-7,8-diol-9,10-epoxide, the ultimate carcinogenic metabolite that binds to DNA.