bluuebunny/crossref_metadata_2025 huggingface dataset:
Cholesterol Risk Management: A Systematic Examination of the Gap from Evidence to Practice
Authors: Olson Kari L. , Bungard Tammy J. , Tsuyuki Ross T.
Published: June 2004

Hypercholesterolemia is a major risk factor for coronary heart disease, and data indicate that aggressive cholesterol reduction decreases mortality and morbidity associated with this disease. Many patients with hypercholesterolemia, however, are not screened, prescribed appropriate lipid‐lowering therapy, or treated to target cholesterol levels. Practice patterns are particularly inadequate for those patients at highest risk for having a cardiac event. We performed a literature search to identify studies of practice patterns in the management of patients with hypercholes‐terolemia with regard to screening, implementing lipid‐lowering therapy, and treating to lipid goals. The findings highlight the potential for substantial opportunities to improve patient outcomes. Future studies should evaluate reasons for suboptimal cholesterol management as well as provide steps to improve management.
Assessment of Hypercholesterolemia Control in a Managed Care Organization
Authors: Straka Robert J. , Taheri Reza , Cooper Susan L. , Tan Agnes W. H. , Smith James C.
Published: June 2004

To determine the extent of achievement of goal low‐density lipoprotein cholesterol (LDL) as defined by National Cholesterol Education Program—Adult Treatment Panel II (NCEP‐ATP II) and American Diabetes Association (ADA) 2000 guidelines, we conducted a retrospective study by integrating data from medical, laboratory, and pharmacy claims databases. Subjects were selected from a 232,000‐member staff‐model managed care organization consisting of 19 clinics in the Minneapolis—St. Paul, Minnesota, metropolitan area. A total of 124,971 members aged 18 years and older, who had been continuously enrolled from July 1, 1996–June 6, 1998, were included. Outcome measures were the extent of achievement of goal LDL as defined by NCEP‐ATP II and the use of antihyperlipidemic drugs for patients with and without diabetes at various levels of risk for coronary heart disease (CHD). Of 124,971 subjects, 6538 had a history of CHD, 1523 of whom met their LDL goal. Of the population with CHD who did not achieve goal, 1141 (43%) missed by over 30 mg/dl; 621 (54%) of these patients were not receiving drug therapy. A total of 17,267 had no history of CHD but had two or more risk factors; 3298 of these achieved their LDL goal. Of those who did not achieve goal, 1136 (35%) missed by over 30 mg/dl; 897 (79%) of these were not receiving drug therapy. A total of 6586 had a history of diabetes; 1004 and 2340 reached an LDL of 100 mg/dl or lower and less than 130 mg/dl, respectively. Of those with diabetes who had an LDL greater than 100 mg/dl, 1276 (49%) missed their goal by over 30 mg/dl; 898 (70%) of these were not receiving drug therapy. Inadequate use of pharmacologic agents plays a significant role in failure to achieve goal LDL for patients with CHD, without CHD, and with diabetes. Analysis of the data based on the new ADA guidelines for LDL demonstrates the need for continued vigilance. Finally, the successful merging of medical, laboratory, and pharmacy claims databases provides a benchmark for other institutions.
Patient‐Reported Adherence to Guidelines of the Sixth Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure
Authors: Cheng Judy W. M. , Kalis Michelle M. , Feifer Stanley
Published: June 2004

Objectives. To compare antihypertensive drug compliance with treatment guidelines established by the Sixth Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI), and to identify patient adherence to antihypertensive drugs and factors affecting prescribing patterns.

Methods. Patients filling antihypertensive drug prescriptions in metropolitan New York area pharmacies were enrolled. Pharmacy externs collected patient‐reported demographics, medical and drug histories, and blood pressure measurements. Compliance with JNC VI guidelines was assessed.

Results. Eight hundred twenty‐one patients from 102 pharmacies participated. Blood pressure was controlled in 61% of patients at the time of the study. The most prescribed class of antihypertensive agents was angiotensin‐converting enzyme inhibitors, followed by diuretics and β‐blockers. Over the study period, compliance with JNC VI guidelines decreased significantly from 85% to 64% (p<0.05). Thirty‐seven percent of patients reported consistent adherence to their antihypertensive regimens. Patients' education level was the only factor found to correlate positively with the appropriateness of antihypertensive agents prescribed.

Conclusion. Compliance with JNC VI guidelines decreased over time, and patient adherence to drug therapy was suboptimal. Continuing‐education efforts to reinforce optimal blood pressure management are necessary.
Retrospective Analysis of Serum Valproate Levels and Need for an Antidepressant Drug
Authors: Stoner Steven C. , Worrel Jodi A. , Vlach David , Jones Michael T. , Ramlatchman Leonard V.
Published: June 2004

We sought to determine whether patients receiving valproate plus an antidepressant had significantly lower serum valproate levels before initiation of the antidepressant than those patients receiving valproate without an antidepressant. We further sought to identify the prevalence of antidepressant‐induced mania and to determine if valproate provided a protective effect against antidepressant‐induced mania. A computer database search from January 1, 1990–June 30, 1998, identified patients with bipolar or schizoaffective disorder treated with valproate. Patients receiving an antidepressant during valproate therapy were identified as the treatment group (9 patients), and the remaining patients served as the control group (17 patients). Serum valproate levels were recorded just before starting the antidepressant for the treatment group and monthly during a comparable period for the control group. The mean time to antidepressant initiation was 15 ± 8 weeks. The mean serum valproate level just before antidepressant initiation was significantly lower for the treatment group compared with the mean serum valproate level averaged over 16 ± 6 weeks for the control group (54 ± 24 vs 73 ± 13 μg/ml, p<0.05). Four patients (44%) developed antidepressant‐induced mania. Three required discontinuation of the antidepressant; their serum valproate levels were 54, 60, and 71 μg/ml. Patients requiring the addition of an antidepressant had significantly lower valproate serum levels than those who did not require an antidepressant. Further study is necessary to determine whether higher serum valproate levels are needed for prevention of depressive symptoms in bipolar and schizoaffective disorders.
Clinical and Microbiological Evaluation of the Periodontal Status of Children with Unilateral Complete Cleft Lip and Palate
Authors: Costa Beatriz , de Oliveira Lima JoséEduardo , Gomide Marcia Ribeiro , Pereira da Silva Rosa Odila
Published: June 2004

Objective

To compare periodontal conditions in children with and without cleft.

Design

Clinical examinations and microbiological analysis of 57 selected children, including 30 with unilateral complete cleft lip and palate (experimental group) and 27 without clefts (control group).

Setting

Hospital of Rehabilitation of Craniofacial Anomalies (HRCA) in Bauru, Sao Paulo, Brazil.

Patients, Participants

All children examined were healthy and between the ages of 5 and 6 years.

Results

The mean plaque index (PI) in the experimental group was higher (1.82 ± 0.3) than in the control group (1.63 ± 0.38), although this difference was not statistically significant. The mean gingival index (GI) in the experimental group (1.82 ± 0.38) was found to be significantly higher (p < .05) than that of the control group (0.79 ± 0.33). The cleft area in the experimental group, with a mean PI of 2.04 ± 0.58 and mean GI of 1.11 ± 0.26, compared with the posterior area, with a mean PI of 1.74 ± 0.37 and mean GI of 1.04 ± 0.26, showed a statistically significant difference only in the PI. Most of the children in both experimental and control groups presented a moderate PI degree (73.33% and 81.48%, respectively) and a high prevalence of mild gingivitis (53.33% and 70.37%, respectively). Analysis of the organisms showed that Prevotella nigrescens was detected in 16.67% of the experimental group and 11.11% of the control, whereas Porphyromonas gingivalis and Treponema denticola were not detected.

Conclusion

Children with clefts showed greater gingival inflammation, despite the same amount of plaque and prevalence of microorganisms.
Growth of Children with Isolated Robin Sequence Treated by Nasopharyngeal Intubation: Importance of a Hypercaloric Diet
Authors: Marques Ilza Lazarini , de Barros Almeida Peres Suely Prieto , Bettiol Heloisa , Barbieri Marco Antonio , Andrea Mauro , de Souza Luiz
Published: June 2004

Objective

To compare the growth curves (weight and length) of two groups of infants with isolated Robin sequence (RS) treated with nasopharyngeal intubation (NPI), one group receiving a hypercaloric diet and the other receiving a normal diet for age, and compare the growth rates and duration of NPI between the two groups.

Design

Prospective longitudinal study.

Setting

Hospital de Reabilitação de Anomalias Craniofaciais, Universidade de São Paulo, Bauru, Brazil, 1998 to 2000.

Patients

Twenty-three children with isolated RS treated by prolonged NPI.

Interventions

Nine children received a milk formula supplemented with 5% to 7% glucose polymers and 3% to 5% medium chain triglycerides (group 1), and 14 children received a milk formula only (group 2). Weight and length were measured at birth and every month up to 6 months of life for the construction of weight and length growth curves. The time of use of the nasogastric tube, age at the beginning of NPI, and duration of NPI were also determined.

Results

No significant difference in length was observed between the two groups. The mean weight curves obtained for the two groups were below the 50th percentile of the National Center for Health Statistics, but weight gain was higher in group 1. The duration of NPI was shorter in this group.

Conclusions

The hypercaloric diet led to an improvement in nutritional status, which in turn contributed to improved respiratory conditions in isolated RS.
Pierre Robin Sequence in Denmark: A Retrospective Population-Based Epidemiological Study
Authors: Printzlau Andreas , Andersen Mikael
Published: June 2004

Objective

To give an epidemiological description of the clinical entity given the name Pierre Robin sequence, defined by retro- and micrognathia, cleft palate, and respiratory distress and describe other malformations and possible intrauterine impairment.

Methods

Using the inclusion criteria of micrognathia, cleft palate, and neonatal respiratory distress, a retrospective population-based study of all Danish live births during 1990 through 1999 were carried out. We found 50 children, 25 boys and 25 girls, fulfilling the inclusion criteria, giving an incidence of 1 in 14,000 live births.

Results

Two-thirds (n = 33) of the children had the classical U-shaped cleft palate. More than one-third (n = 19) had one or several other malformations, and in five patients the triad of Pierre Robin was a minor feature of a complex syndrome. The most common noncomplex syndrome was the Stickler syndrome found in 6 of the 50 patients. More than one-fourth (n = 17) had some kind of intrauterine impairment, with no specific obstetric diagnosis predominant. Consistent with this, the average birth weight was well below normal.

Conclusions

Several authors have stated that the triad of Pierre Robin is not a nosological entity, in that it has diverse etiology and diverse pathogenesis. We conclude that the triad of Pierre Robin still can be regarded as a clinical entity, readily defined at birth, experiencing the same neonatal problems in varying degrees and hence the possibility of designing treatment protocols for later scientific evaluation.
Cost-Effectiveness of Infant Orthopedic Treatment regarding Speech in Patients with Complete Unilateral Cleft Lip and Palate: A Randomized Three-Center Trial in the Netherlands (Dutchcleft)
Authors: Konst Emmy M. , Prahl Charlotte , Weersink-Braks Hanny , De Boo Theo , Prahl-Andersen Birte , Kuijpers-Jagtman Anne M. , Severens Johan L.
Published: June 2004

Objective

To investigate the cost-effectiveness of infant orthopedic treatment (IO), compared with no such treatment in children with complete unilateral cleft lip and palate (UCLP) focusing on the effects on speech development at age 2.5 years.

Design

In a three-center prospective, randomized clinical trial (Dutchcleft), two groups of children with complete UCLP were followed longitudinally: one group was treated with IO based on a modified Zurich approach in the first year of life (IO group) and the other group did not receive this treatment (non-IO group).

Patients

The participants had complete UCLP without soft tissue bands or other malformations. They were born at term and their parents were native Dutch speakers.

Outcome Measures

The effect of IO on speech development at age 2.5 years was measured blindly by five expert listeners judging the “total impression of speech quality” on a 10-point equal-appearing interval scale. Costs were measured from a societal viewpoint in Euro.

Results and Conclusion

The IO group (n = 10) obtained a statistically significant higher rating, compared with the non-IO group (n = 10). The effect size was large, indicating that the improvement may be considered a clinically important change. The cost for treatment by the orthodontist was higher in the IO group. For both groups, the mean cost was related to the mean rating for “total impression of speech quality.” The resulting cost-effectiveness for IO, compared to non-IO was €1041 for 1.34 point speech quality improvement. The financial investment that is necessary to obtain this improvement seems limited. Thus, from the perspective of speech development, the cost-effectiveness of IO over non-IO seems acceptable at this point in time.
Alveolar Ridge Notching as a Predictor for Secondary Bone Grafting in Incomplete Alveolar Clefts
Authors: Kita Hiroki , Kochi Shoko , Kamiya Noriaki , Yamada Atsushi , Mitani Hideo
Published: June 2004

Objectives

To clarify the need for secondary alveolar bone grafting (ABG) in incomplete alveolar clefts identified clinically from its appearance and test the usefulness of alveolar ridge notching in the edentulous stage as a predictor for ABG.

Design

A prospective, cross-sectional study comparing three groups of patients at two stages.

Patients

Twenty-four patients with unilateral incomplete cleft of primary palate who underwent either no surgery or cheiloplasty only.

Methods

Cleft severity was assessed at two stages. Initially, the appearance of untreated clefts was assessed before cheiloplasty or at 3 months of age in the patient without cheiloplasty and graded by severity into three groups. Later alveolar bone defects were assessed with computed tomography in the primary or mixed dentition period. A decision regarding the requirement for ABG was then made. Finally, the relationship between untreated clefts and the need for ABG was determined.

Main Outcome Measures

Nine (75%) of 12 patients with alveolar ridge notching in the edentulous stage were diagnosed as fulfilling the criteria for ABG.

Results

The alveolar bone defects correlated with the severity of untreated cleft conditions (Spearman r = 0.65, p = .002). The need for ABG was greater in patients with alveolar ridge notching, compared with those without notching (9/12 versus 0/12, p = .00034).

Conclusions

Most patients with incomplete alveolar clefts clinically identified in the edentulous stage required ABG, and alveolar ridge notching in the edentulous stage could therefore serve as a predictor for ABG.
Choanal Atresia and Rare Craniofacial Clefts: Report of Three Cases with a Review of the Literature
Authors: Bonafos G. , Capon-Degardin N. , Fayoux P. , Pellerin P.
Published: June 2004

Objective

This retrospective study reports three cases associating choanal atresia (CA) and rare craniofacial clefts (RCFCs) described in Tessier's classification. Although still discussed, these two malformations could derive from the same embryopathogenic context, with an anomaly of migration of the neural crest cells as a common factor. In two of the three cases presented, the RCFC was diagnosed during the neonatal period, and the third case (case 3) was diagnosed at the age of 12 years. The knowledge of this malformative association must permit the diagnosis of CA as early as the neonatal period or through the thorough and directed examination of the children as part of the follow-up.
The Buccal Flap—A Useful Technique in Cleft Palate Repair?
Authors: Jackson Ian T. , Moreira-Gonzalez Andrea A. , Rogers Amy , Beal Barbara J.
Published: June 2004

Objective

To evaluate speech quality and oronasal fistula after primary palate repair using a buccal mucosal flap.

Design

Retrospective study cohort of patients with cleft palate.

Setting

Primary care center for treatment of craniofacial congenital anomalies.

Patients and Method

One hundred fifty-six nonsyndromic patients underwent palatoplasty with the buccal myomucosal flap by the senior surgeon between 1989 and 2002. The preoperative workup, surgical technique, and other factors that might affect the outcome were identical in every case. Oronasal fistula and variables affecting speech quality were analyzed.

Results

The most common type of cleft was unilateral cleft lip and palate (43.5%). The median follow-up was 5.8 years (0.4 to 21 years), and the median age at repair was 6.2 months. The overall fistula formation was 3.6%, decreasing progressively: 1989 to 1994: 2.9%, 1995 to 2002: 0.7% (p < .05). Velopharyngeal incompetence (VPI) occurred in 8.8% of the patients, decreasing from 5.3% to 3.5% in the last years. VPI and oronasal fistulae were observed mainly in unilateral and bilateral clefts of the lip and palate. Velopharyngeal adequacy occurred in 91.1% of the children, and resonance was normal in 91.1 %. None of the patients had severe hypernasality or hyponasality. Articulation was normal in 97.9% of the children. Speech quality was good in 89% of the patients.

Conclusions

The technique presented has been effective, with the advantages of palatal closure without tension, good muscular reconstruction, lengthening of the nasal layer, and palatal closure without raw areas. The technique, early repair, and surgeon's skills were the most important variables for good outcomes regarding speech and fistula formation.
Correction of Facial Asymmetry with Multiplanar Mandible Distraction: A One-Year Follow-Up Study
Authors: Ko Ellen W. , Hung Kai-Fong , Huang Chiung Shing , Chen Philip K. T.
Published: June 2004

Objective

To assess the effectiveness of multiplanar mandibular distraction by evaluating the treatment effect and 1-year stability, measuring changes on the affected and nonaffected sides, and evaluating correction of the occlusal plane and oral commissure cant.

Patients and Method

Eleven patients aged 5 to 9.4 years (mean 6.7 years) who underwent unilateral multiplanar mandibular distraction were included, 10 with hemifacial microsomia and one temporomandibular joint ankylosis. Intermaxillary elastics were applied to intraoral dental devices during and after distraction until bony consolidation and occlusal interdigitation were achieved. Radiographs were taken and measured before distraction, after vertical distraction, after completion of distraction, and 1 year after treatment. Frontal facial photographs were obtained to analyze the changes in the position of the oral commissure.

Results

The facial profile was improved by sagittal mandibular advancement. Although the facial height increased 6.6 mm during vertical distraction, with a net gain of 5.8 mm in follow-up, the mandibular plane angle was maintained. The affected mandibular length increased 8.3 mm after distraction and remained unchanged after 1 year. Ramus height increased 12.7 mm after distraction and relapsed 3.8 mm at follow-up (30%). The affected body length demonstrated postoperative growth of 3.1 mm. Correction of the chin deviation was 9.8 mm after distraction and relapsed 1.6 mm (16.3%) after 1 year. Canting of the occlusal plane and oral commissure was corrected and remained stable.

Conclusion

Multiplanar mandibular distraction is an effective treatment for correcting facial asymmetry in young children. The 1-year follow-up revealed that the new sagittal jaw relation and mandibular body length were stable, and the achieved occlusal interdigitation was well maintained. However, the ramus height and chin deviation demonstrated some relapse.
A Case of Oculo-Facio-Cardio-Dental Syndrome with Integrated Orthodontic-Prosthodontic Treatment
Authors: Kawamoto Tatsuo , Motohashi Nobuyoshi , Ohyama Kimie
Published: June 2004

Objective

Oculo-facio-cardio-dental (OFCD) syndrome is a very rare condition that requires comprehensive dental management because of associated multiple dental anomalies such as canine radiculomegaly, delayed dentition, oligodontia, persistent primary teeth, microdontia, and macrodontia. This report presents a case of OFCD in a Japanese girl (13 years 1 month old). We analyzed cephalograms, panoramic roentgenograms, and dental casts and discuss our integrated orthodontic-prosthodontic treatment.

Design

The sizes of the tooth crown and root as well as lateral cephalograms were compared with those from a Japanese control group. The outcome of orthodontic treatment was evaluated by comparing cephalograms taken before and after treatment.

Results

Radiculomegaly was found not only in the upper and lower canines but also the upper central incisors and lower first premolars. Macrodontia was found in the upper central incisors, upper canines, lower canines, and lower first premolars. Microdontia was found in the upper lateral incisor. Lateral cephalometric analysis showed a remarkable hypoplastic midface in both the sagittal and vertical dimensions, coupled with a significantly decreased cranial base length and an increased gonial angle. Serial lateral cephalograms during orthodontic treatment from 13 to 23 years of age demonstrated only slight maxillary growth and significant downward mandibular growth with clockwise rotation in addition to pronounced labial tipping of the upper central incisors.

Conclusions

Characteristic dental anomalies together with a unique craniofacial dysmorphology were clarified. Successful oral rehabilitation was achieved by integrated orthodontic-prosthodontic treatment.
Methodology for Perceptual Assessment of Speech in Patients with Cleft Palate: A Critical Review of the Literature
Authors: Lohmander Anette , Olsson Maria
Published: June 2004

Objective and Design

This review of 88 articles in three international journals was undertaken for the purpose of investigating the methodology for perceptual speech assessment in patients with cleft palate. The articles were published between 1980 and 2000 in the Cleft Palate–Craniofacial Journal, the International Journal of Language and Communication Disorders, and Folia Phoniatrica et Logopaedica.

Results and Conclusions

The majority of articles (76) were published in the Cleft Palate–Craniofacial Journal, with an increase in articles during the 1990s and 2000. Information about measures or variables was clearly given in all articles. However, the review raises several major concerns regarding method for collection and documentation of data and method for measurement. The most distressing findings were the use of a cross-sectional design in studies of few patients with large age ranges and different types of clefts, the use of highly variable speech samples, and the lack of information about listeners and on reliability. It is hoped that ongoing national and international collaborative efforts to standardize procedures for collection and analysis of perceptual data will help to eliminate such concerns and thus make comparison of published results possible in the future.
Association between 10 Microsatellite Markers and Nonsyndromic Cleft Lip Palate in the Chilean Population
Authors: Blanco Rafael , Suazo José , Santos JoséLuis , Paredes Mónica , Sung Hsiao , Carreño Hernán , Jara Lilian
Published: June 2004

Objective

The objective of this case-control study was to evaluate the possible association between nonsyndromic cleft lip/palate (NSCLP) and 10 genetic markers in four chromosomal regions in the admixed Spanish-Amerindian Chilean population.

Setting

Study participants included 56 patients with NSCLP identified and interviewed for positive family history during the course of clinical examinations at different rehabilitation centers in the cities of Santiago and Talca, Chile. A control group of 59 normal individuals without known familial antecedents of clefting was obtained from blood bank donors of the University Hospital, University of Chile. Cases and controls belonged to low- to low-middle socioeconomic strata.

Results

Ten markers from chromosome 4p, 4q, 6p, 17q, and 19q were assessed (MSX1, D4S175, D4S192, F13A1, EDN1, D6S89, D6S105, D6S109, D17S579, BCL3). Four of them showed significant deviations from Hardy-Weinberg expectations in controls, according to the exact test (D4S192, BCL3, F13A1, and D6S89). The case-control comparison by means of the CLUMP program showed significant differences only in BCL3, and D6S109 almost reached statistical significance.

Conclusions

Most of the genetic regions with positive results in Caucasian populations may not be involved in NSCLP in Chile, regardless of the positive evidence for the candidate region on chromosome 19. Similar findings have been reported recently in the Chinese population.
Comparison of Obstructive Sleep Apnea Syndrome in Children with Cleft Palate following Furlow Palatoplasty or Pharyngeal Flap for Velopharyngeal Insufficiency
Authors: Liao Yu-Fang , Noordhoff M. Samuel , Huang Chiung-Shing , Chen Philip K. T. , Chen Ning-Hung , Yun Claudia , Chuang Ming-Lung
Published: June 2004

Objective

To evaluate the incidence and severity of obstructive sleep apnea syndrome (OSAS) in patients with cleft palate having a Furlow palatoplasty or pharyngeal flap for correction of velopharyngeal insufficiency (VPI).

Patients

A total of 48 nonsyndromic children with repaired cleft palate with VPI were enrolled in the study. Twenty of the children had a Furlow palatoplasty (F group) and 28 children had a pharyngeal flap (P group) for correction of VPI.

Interventions

An overnight polysomnography evaluation was done to evaluate the incidence and severity of OSAS 6 months or more postoperatively.

Main Outcome Measures

Symptoms of OSAS, respiratory disturbance index (RDI), oxyhemoglobin desaturation index (DI), and sleep stages were measured.

Results

In the P group, the mean percentage of stage 2 sleep was lower than the F group (p < .05). The mean RDI and DI were larger in the P group, compared with the F group (p < .001). The incidence and severity of OSAS were higher in the P group, compared with the F group (p < .001 and p = 0.05, respectively).

Conclusions

A Furlow palatoplasty should be used in deference to a pharyngeal flap whenever possible on the basis of the preoperative evaluation of VPI because of the decreased incidence and severity of OSAS.
Cleft Palate and Congenital Synechiae Syndrome: A Case Report
Authors: Murphy Siun M. , Rea Suzanne , McGovern Eleanor , Fleming Paddy , Orr David
Published: June 2004

Objective

A 1-day-old baby girl with a cleft of the secondary palate and a soft tissue band connecting the upper and lower jaws and preventing mouth opening was referred to the cleft lip and palate team by her pediatrician. This case represents a further example of an interesting but rare anomaly known as congenital alveolar synechia syndrome that requires early management to allow normal feeding and oral development.
Stabilization of an Edentulous Premaxilla for an Alveolar Bone Graft: Case Report
Authors: Bohman Paul , Yamashita Dennis Duke , Baek Seung-Hak , Yen Stephen L.-K.
Published: June 2004

Objective

The patient with bilateral cleft lip and palate and an edentulous premaxilla poses several additional challenges to the surgeon and the orthodontist. The cleft segments are usually expanded and stabilized with tooth-supported orthodontic appliances prior to an alveolar bone graft. This report describes a technique for stabilizing an edentulous premaxilla using an orthodontic-surgical screw that is ligated to an orthodontic archwire.
Current Concepts in Cell Toxicity
Authors: Samali Afshin , Zhivotovsky Boris , Orrenius Sten
Published: June 2004

This overview provides a basic definition of cellular death and the mechanisms that are associated with cell death: apoptosis and necrosis. It includes a description of structural changes and macromolecular degradation and the roles of signaling, mitochondria, and genetic changes in the two forms of cell death.
In Situ Hybridization Histochemistry
Authors: Kokaia Zaal
Published: June 2004

This unit describes two methods of in situ hybridization: one uses an 35S‐labeled oligonucleotide probe and the other uses a digoxigenin‐labeled oligonucleotide probe on frozen, cryostat‐sectioned samples. These methods allow detection of the physical distribution and expression levels of target mRNA. Protocols are also included for labeling the probes and preparing the sample material.
The Salmonella (Ames) Test for Mutagenicity
Authors: Zeiger Errol , Mortelmans Kristien
Published: June 2004

The Ames test is a widely accepted short‐term assay for detecting chemicals that induce mutations in the DNA of organisms. The assay is a reverse mutation assay that detects the mutational reversion of his‐dependent Salmonella to his‐independent colonies. Mutation‐causing chemicals increase the frequency of occurrence of his‐independent colonies, usually in a dose‐dependent fashion. This unit includes protocols for the basic plate test with and without preincubation as well as procedures for highly volatile chemicals and gases, chemicals that require reductive metabolism, and chemicals that are available in small quantities. In addition there are protocols for dose selection, strain maintenance and preservation, freezing working and permanent cultures, and preparing the S‐9 metabolic activation system.
Measurement of a Malondialdehyde‐DNA Adduct
Authors: Plastaras John P. , Marnett Lawrence J.
Published: June 2004

Determining the levels of various DNA adducts has become an essential tool in understanding the toxicology of carcinogens. Direct measurement of DNA adduct levels, the true biologically effective dose of a mutagen, can be correlated with biological outcomes or used to probe mechanisms of adduct formation. Each adduct to be measured requires a specific assay. Malondialdehyde is a carcinogenic and mutagenic electrophile that is endogenously produced during peroxidation of polyunsaturated fatty acids. Its reaction with deoxyguanosine produces a fluorescent exocyclic pyrimidopurinone that can be detected by gas chromatographic/negative chemical ionization‐electron capture mass spectroscopy. Methods for preparing an immunoaffinity gel and for HPLC quatification of nucleosides are also included.
Assays for DNA Damage
Authors: Sutherland Jessica E. , Costa Max
Published: June 2004

This unit describes several assays for detecting several kinds of DNA damage (strand breaks, internal crosslinking, DNA/protein crosslinks) and repair activity following exposure to genotoxic agents. The methods include single‐cell electrophoresis (comet assay), filter eluting, K‐SDS precipitation, and measurement of unscheduled DNA synthesis.
Assays for Detecting Chromosomal Aberrations
Authors: Klein Catherine B. , Broday Limor , Costa Max
Published: June 2004

This unit describes the techniques used to investigate chromosome structure to detect changes induced by toxicant exposure. Protocols are included for detecting microcucleus (MN) formation, changes in banding patterns by Giemsa banding or fluorescence in situ hybridization (FISH) using chromosome‐specific probes, and sister chromatid exchange (SCE) in cultures of established cell lines.
Methods for Measuring DNA Adducts and Abasic Sites I: Isolation, Purification, and Analysis of DNA Adducts in Intact DNA
Authors: Swenberg James A. , Ham Amy‐Joan L. , McDorman Kevin S. , Morinello Eric J. , Nakamura Jun , Schoonhoven Robert
Published: June 2004

The major event involved in the formation of mutations and the initiation and progression of cancer is the induction of DNA damage by reactive intermediates arising from exposure to endogenous and exogenous chemicals. Many electrophilic metabolites of chemicals covalently bind to the bases of DNA causing specific DNA adducts. This unit includes protocols for preparing samples of intact DNA and adduct analysis to quantify the number of adducts that can potentially cause mutagenic or carcinogenic damage.
Methods for Measuring DNA Adducts and Abasic Sites II: Methods for Measurement of DNA Adducts
Authors: Swenberg James A. , Ham Amy‐Joan L. , Koc Hasan , La David K. , Morinello Eric J. , Pachkowski Brian F. , Ranasinghe Asoka , Upton Patricia B.
Published: June 2004

This unit contains protocols for analyzing DNA adducts separated from the DNA backbone. HPLC is used to quantify total guanine or ribo‐ or deoxynucleotides as well as methods for analyzing specific adducts. These methods include HPLC with electrochemical detection, immunoaffininty chromatography to enrich for specific adducts, and gas and liquid chromatography in combination with HPLC and mass spectrometry.
Measurement of Cytochrome P‐450
Authors: Schenkman John B. , Jansson Ingela
Published: June 2004

The family of cytochrome P‐450 enzymes are involved in the metabolism of xenobiotics. This unit provides protocols for the three most common ways to measure cytochrome P‐450. Spectrophotometric detection of the Soret band gives a quantitative measure of total cytochrome P‐450 in microsomes, liver homogenate, or mitochondria, but lacks information about the specific P‐450 forms. Immunochemical methods such as immunoblotting provide very sensitive semiquantitative identification of specific P‐450 enzymes Finally, enzymatic assays depend on substrate specificity and also provide information about specific P‐450 enzymes, although some members of this group of enzymes have broad and sometimes overlapping specificities.
Purification of Cytochrome P‐450 Enzymes
Authors: Bell‐Parikh L.C. , Hosea N.A. , Martin M.V. , Guengerich F.P.
Published: June 2004

Among the liver P‐450 xenobiotic‐metabolizing enzymes, P450‐2E1 is of interest because of its activation of potent carcinogens, and P‐450 1A2 is of interest because of its role in oxidation of drugs and carcinogens. This unit describes column chromatography protocols for purification of recombinant forms of these enzymes expressed in a bacterial expression system.
Measurement of Aryl and Alcohol Sulfotransferase Activity
Authors: Sheng Jonathan J. , Sharma Vyas , Duffel Michael W.
Published: June 2004

Two methods are presented for determining the catalytic activity of aryl and alcohol sulfotransferases. The first assay is a simple and rapid procedure that is based on the extraction of a paired ion formed between the product organic sulfate and methylene blue. The second method employs HPLC analysis of the substrate‐dependent formation of PAP, an assay that is particularly useful when the sulfuric acid ester product is chemically unstable.
Measurements of Flavin‐Containing Monooxygenase (FMO) Activities
Authors: Rose Randy L.
Published: June 2004

Measurement of Flavin‐Containing Monooxygenase (FMO) Activities (Randy L. Rose, North Carolina State University, Raleigh, North Carolina). This unit describes methods used for measuring the presence of flavin‐containing monooxygenases using NADPH oxygenation and methamizole oxidation. Methods are also provided to determine the relative contributions of FMO versus cytochrome P450 from microsomes.
Techniques for Measuring the Activity of Carboxylic Acid:CoA Ligase and Acyl‐CoA:Amino Acid N‐Acyltransferase: The Amino Acid Conjugation Pathway
Authors: Kelley Michael , Vessey Donald A.
Published: June 2004

A wide variety of xenobiotic carboxylic acids are metabolized to their amino acid conjugates via a pathway that exists primarily in liver and kidney. This conjugation occurs in a two‐step pathway catalyzed by two distince types of enzymes, ligases and transferases. Measurements of acyl‐CoA ligase activity include monitoring the rate of appearance of AMP or PPi, or the CoA adduct. N‐acyltransferases catalyze formation of an amino acid conjugate from the CoA‐activated intermediate, releasing CoA. This reaction is monitored by following the release of free CoA or the disappearance of the acyl‐CoA adduct.
Determination of Paraoxonase 1 Status and Genotypes at Specific Polymorphic Sites
Authors: Richter Rebecca J. , Jampsa Rachel L. , Jarvik Gail P. , Costa Lucio G. , Furlong Clement E.
Published: June 2004

The procedures for determining paraoxonase (PON1) status and for determining PON1 genotypes for polymorphisms in coding and important regulatory regions are described. PON1 status is determined by a functional two‐substrate analysis of plasma PON1 activities. Differences in catalytic efficiency of the PON1192Q and PON1192R alloforms result in the clear separation of all three phenotypes at position 192 (Q/Q, Q/R, R/R) and at the same time, the two‐substrate analysis indicates activity levels of PON1. Because the enzyme activity levels are as important as the polymorphic genotypes, this two‐substrate analysis of PON1 status provides the most relevant information for investigating the association of PON1 genetics with susceptibilities to disease, organophosphorus insecticide sensitivity, and pharmacokinetic status of drug metabolism. Genotyping of polymorphic sites alone fails to provide this important information but can be useful for gene frequency determination and forensic analysis. Analytical procedures for determining PON1 status and genotypes are described.
Measurement of Bioavailability: Measuring Absorption Through Skin In Vivo in Rats and Humans
Authors: Bounds Stephen V. , Hawkins David R.
Published: June 2004

Absorption through the skin is a route of exposure to a wide variety of therapeutic and/or environmental compounds. In vivo assays are advantageous in that they retain intact epidermal and dermal structures and thus reflect a more normal situation. The test compound is applied to the skin in a protected area for a specified period of time. At the end of the incubation, skin, tissues, and excreta are assessed for the presence of the test compound. The assay can also be performed with volatile compounds. When performed using laboratory animals, it is possible to assess the distribution of the compound throughout the body. If human volunteers are studied such analysis is limited to the skin, blood, and excreta.
Measurement of Disposition Half‐Life, Clearance, and Residence Times
Authors: Mayersohn Michael
Published: June 2004

This unit provides an overview of the principles underlying the elimination process. Experimental design issues, methods of data analysis, and complications that may be encountered are discussed. A complete worked example is provided to exemplify the methods of analysis that are available.
Measurement of Glutathione and Glutathione Disulfide
Authors: Mustacich Debbie
Published: June 2004

Measurements of glutathione should include quantification of both the reduced and oxidized forms. HPLC‐based assays for glutathione and other cellular thiols and disulfides utilize a variety of detection methods, including ultraviolet, fluorescence, and electrochemical methods. This unit provides rapid, sensitive, highly versatile and reproducible HPLC methods for separating and quantifying sulfur‐containing amino acids and related derivatives from a variety of biological samples.
HPLC‐Based Assays for Enzymes of Glutathione Biosynthesis
Authors: White Collin C. , Krejsa Cecile J. , Eaton David L. , Kavanagh Terrance J.
Published: June 2004

Glutamate cysteine ligase and gluthione synthase carry out the two‐step synthesis of glutathione. The fluorescent thiol‐reactive compound monobromobimane is used to derivatize reaction products in an HPLC‐based assay with fluorescence detection. The assay described in this unit can be adapted for tissue homogenates or cultured cells.
γ‐Glutamyl Transpeptidase Activity Assay
Authors: Ikeda Yoshitaka , Taniguchi Naoyuki
Published: June 2004

Gamma‐glutamyl transpeptidase catalyzes the transfer of a gamma‐glutamyl bond and is of importance in glutathione metabolism. The most common assay methods for this enzyme, in which gamma‐glutamyl derivatives of p‐nitroanilineare used as a gamma‐glutamyl donor, are described in this unit.
Oxidant‐Induced Regulation of Glutathione Synthesis
Authors: Liu Rui‐Ming , Choi Jinah , Forman Henry J.
Published: June 2004

This unit describes protocols for characterizing the expression of two glutathione biosynthesis enzymes: gamma‐glutamylcysteine synthase (GCS) and gamma‐glutamyl transpeptidase (GGT) in response to oxidants. GCS catalyzes the first and rate‐limiting step of glutathione synthesis, while GGT degrades extracellular glutathione (GSH) to provide the amino acids required for intracellular synthesis of GSH. Northern blot hybridization is used to quantitatively assess the mRNAs for the two enzymes in response to oxidant. Nuclear run‐on is used to determine the rate of transcription.
Measurement of Thioredoxin and Thioredoxin Reductase
Authors: Arnér Elias S.J. , Holmgren Arne
Published: June 2004

The thioredoxin system is ubiquitous, providing reducing equivalents to essential biosynthetic enzymes like ribonucleotide reductase. It is essential for cellular redox regulation, control of oxidative stress, and protection against oxidative damage. This unit includes protocols for measuring thioredoxin or thioredoxin reductase in biological preparations or as purified enzymes.
The Heme Biosynthesis Pathway and Clinical Manifestations of Abnormal Function
Authors: Phillips John D. , Kushner James P.
Published: June 2004

Biosynthesis of heme is important for both prokaryotes and eukaryotes. The enzymes for this multistep process are distributed between the cytosol and mitochondria in eukaryotes. In humans there are inherited and acquired disorders characterized by over synthesis of one or more enzymes or absence of an enzyme. This overview discusses each enzyme in the heme biosynthesis pathway.
Measurement of Heme Concentration
Authors: Sinclair Peter R. , Gorman Nadia , Jacobs Judith M.
Published: June 2004

Heme (iron protoporphyrin IX) is a prosthetic group for a number of hemoproteins in different tissues (e.g., hemoglobin, myoglobin, cytochrome P‐450s, mitochondrial cytochromes, catalases, and peroxidases). Mutations in the biosynthetic pathway can affect the synthesis and/or degradation of heme. Several assays are provided in this unit for quantifying heme: a spectophotometric assay based on the characteristic absorption spectrum of oxidized and reduced form of the hemochrome formed by replacing the nitrogen ligands with pyridine; a fluorescence assay based on removal of the iron by a heated, strong oxalic acid solution to produce fluorescent protoporphyrin; a reversed‐phase HPLC assay to measure heme and intermediates in the synthetic pathway; and a radiometric assay to measure newly synthesized heme in tissue culture cells.
Measurement of δ‐Aminolevulinate Dehydratase Activity
Authors: Fujita Hiroyoshi
Published: June 2004

Vertebrate ALAD is a cytosolic enzyme that catalyzes the second step of porphyrin formation, and it is expressed abundantly in liver and erythroid tissues as well as in other tissues. This unit describes a colorimetric assay for measuring ALAD activity in mammalian blood or tissues and the extent of reactivation of the enzyme in lead‐inhibited samples.
Measurement of Ferrochelatase Activity
Authors: Taketani Shigeru
Published: June 2004

Ferrochelatase is the terminal enzyme in the heme biosynthesis pathway. Under anaerobic conditions it catalyzes the insertion of ferrous iron into the protoporphyrin IX ring to form protoheme. In the absence of iron and under aerobic conditions, the enzyme will use zinc or mercury as a substitute. The assay described in this unit uses zinc under aerobic conditions and the artificial substrate mesoporphyrin to analyze the formation of zinc‐mesoporphyrin by HPLC with UV or fluorescence detection.
Measurement of Erythrocyte Protoporphyrin Concentration by Double Extraction and Spectrofluorometry
Authors: Parsons Patrick J.
Published: June 2004

Quantification of the level of erythrocyte protoporphyrin can be used as a screening assay for lead exposure. In this method, porphyrins and heme compounds are extracted from whole blood using ethyl acetate, and porphyrins are then separated from heme by back‐extraction with HCl solution. The extracted porphyrins are then quantified using a spectrofluorometer calibrated with protoporphyrin IX standard solutions.
Detection of Heme Oxygenase Activity by Measurement of CO
Authors: Vreman Hendrik J. , Stevenson David K.
Published: June 2004

Heme oxygenase (HO) is the first and rate‐limiting step in degradation of heme, and in the presence of NADPH‐cytochrome P‐450 reductase it produces equimolar amounts of biliverdin and CO. CO produced in a closed system can be quantified as described in this unit by gas chromatography as a measure of HO activity in tissue slices, tissue homogenates, and tissue fractions.
Histochemical Analysis of Heme Degradation Enzymes
Authors: Ewing James F.
Published: June 2004

Histochemical analysis can be used to study the cellular localization, expression, and regulation of enzymes responsible for catabolism of the heme molecule. This unit describes protocols for assessing the expression and regulation of specific HO‐1 and HO‐2 mRNAs by in situ hybridization using digoxigenin‐tagged probes. Immunohistochemistry is used to visualize specific cellular sites of HO‐1 and HO‐2 protein expression.
Quantitation of Human Heme Oxygenase (HO‐1) Copies by Competitive RT‐PCR
Authors: Abraham N.G. , Shenouda S.K. , Goodman A.
Published: June 2004

The authors have developed a system for quantifying human HO‐1 mRNA in samples limited in cell number and/or mRNA copies. Total RNA from human liver was used to develop the system. The RNA is reverse transcribed and amplified by PCR in a tube also containing an internal standard obtained by deleting 50 base pairs from the original human gene. After amplification the two templates are resolved and quantified. When the internal standard is present in the reaction mixture, the ratio of amplified sample to internal standard is proportional to the amount of sample RNA making it possible to calculate the number of specific mRNA molecules.
Purification and Characterization of Heme Oxygenase
Authors: Wilks Angela
Published: June 2004

High‐yield expression and purification of human heme oxygenase isozyme 1 provided the breakthrough in characterizing the protein from mechanistic and structural standpoints. This unit provides a protocol for high‐level expression and subsequent purification of HO‐1. The commentary includes a discussion of subsequent biochemical and biophysical characterizations.
Assay of Tissue Activity of Nitric Oxide Synthase
Authors: Schmidt Kurt , Mayer Bernd
Published: June 2004

Nitric oxide synthase activity can be assayed by a variety of methods. The simplest method to assay activity is the conversion of radiolabeled L‐arginine to L‐citrulline. The assay described in this unit is sensitive, simple to perform, is relatively robust, and can be performed with intact monolayer or suspension cells, or with cell or tissue extracts. The product is easily separated from the reaction by cation‐exchange chromatography.