Efficacy of Eating Disorder Focused Family Therapy for Adolescents With Anorexia Nervosa: A Systematic Review and Meta-Analysis.

OBJECTIVE: To systematically review and evaluate the efficacy of eating disorder focused family therapy (FT-ED) in comparison to all other forms of psychotherapy for children and adolescents with anorexia nervosa. A secondary aim is to assess the relative efficacy of different variations of FT-ED (e.g., shorter vs. longer dose, parent-focused). METHODS: A search with relevant terms was systematically conducted on four databases. Twenty-three publications across 18 randomized controlled trials met inclusion criteria. Outcomes of interest included variables related to weight, eating psychopathology, and remission status. Study quality was assessed, and data were extracted by two independent researchers. RESULTS: Adolescents receiving FT-ED gained significantly more weight by the end of treatment in comparison to those receiving individual psychotherapy. FT-ED that was delivered just to parents or to parents and child separately offered preferable weight outcomes and rates of recovery at the end of treatment in comparison to conjoint FT-ED. No other outcomes tested in the meta-analysis were statistically significant at the end of treatment or follow-up. DISCUSSION: Currently available data suggest the use of FT-ED in its conjoint or separated/parent focused format is the best outpatient treatment option for adolescents with anorexia nervosa when immediate weight gain is paramount. The variability of outcome measurement, including the tools used and timepoints chosen, limit comparison among no more than a handful of studies. The field would benefit from the standardization of measurement and reporting guidelines for future clinical trials. TRIAL REGISTRATION: PROSPERO number: CRD42023396263.

OBJETIVO: Revisar y evaluar sistemáticamente la eficacia de la terapia familiar centrada en el trastorno de conducta alimentaria (TF­TCA; FT­ED por sus siglas en inglés) en comparación con todas las demás formas de psicoterapia para niños y adolescentes que padecen anorexia nerviosa. Un objetivo secundario es evaluar la eficacia relativa de diferentes variaciones de la TF­TCA (por ejemplo, dosis más corta vs. más larga, centrada en los padres). MÉTODOS: Se realizó una búsqueda sistemática con términos relevantes en cuatro bases de datos. Veintitrés publicaciones de 18 ensayos controlados aleatorios cumplieron con los criterios de inclusión. Los resultados de interés incluyeron variables relacionadas con el peso, la psicopatología alimentaria y el estado de remisión. La calidad del estudio fue evaluada y los datos fueron extraídos por dos investigadores independientes. RESULTADOS: Los adolescentes que recibieron TF­TCA ganaron significativamente más peso al final del tratamiento en comparación con aquellos que recibieron psicoterapia individual. La TF­TCA que se administró solo a los padres o a padres e hijos por separado ofreció mejores resultados en el peso y tasas de recuperación al final del tratamiento en comparación con la TF­TCA conjunta. Ningún otro resultado probado en el metaanálisis fue estadísticamente significativo al final del tratamiento o durante el seguimiento. DISCUSIÓN: Los datos disponibles actualmente sugieren que el uso de la TF­TCA en su formato conjunto o separado/centrado en los padres es la mejor opción de tratamiento ambulatorio para adolescentes que padecen anorexia nerviosa cuando la ganancia de peso inmediata es primordial. La variabilidad en la medición de los resultados, incluyendo las herramientas utilizadas y los puntos temporales elegidos, limita la comparación entre no más de un puñado de estudios. El campo se beneficiaría de la estandarización de la medición y las directrices de reporte para futuros ensayos clínicos.

Recommendations on breast cancer screening and prevention in the context of implementing risk stratification: impending changes to current policies.

In recent years, risk stratification has sparked interest as an innovative approach to disease screening and prevention. The approach effectively personalizes individual risk, opening the way to screening and prevention interventions that are adapted to subpopulations. The international perspective project, which is developing risk stratification for breast cancer, aims to support the integration of its screening approach into clinical practice through comprehensive tool-building. Policies and guidelines for risk stratification-unlike those for population screening programs, which are currently well regulated-are still under development. Indeed, the development of guidelines for risk stratification reflects the translational aspects of perspective. Here, we describe the risk stratification process that was devised in the context of perspective, and we then explain the consensus-based method used to develop recommendations for breast cancer screening and prevention in a risk-stratification approach. Lastly, we discuss how the recommendations might affect current screening policies.

Construct validity and reliability of a French-Canadian translation of the eating in the absence of hunger questionnaire for children and adolescents.

Eating in the absence of hunger (EAH) has been associated with overweight and obesity during childhood. The gold standard to assess this behavior is a laboratory-based protocol, but a questionnaire to assess EAH more efficiently in children and adolescents has been developed and validated in English. We assessed construct validity (structural and convergent validity) and reliability (internal consistency and temporal stability) of a French translation of the EAH Questionnaire for Children and Adolescents among French-Canadian youths. We recruited participants in Montreal (Canada) aged 7-15 years old, who completed the questionnaire and provided anthropometric data. We asked participants to complete the questionnaire a second time ∼4 weeks later. The questionnaire consists of 14 questions and 3 subscales that assess EAH due to negative affect, fatigue/boredom, and external cues. We performed an exploratory factor analysis to test the factor structure and we calculated Cronbach alpha coefficients and intra-class correlations to assess internal consistency and temporal stability, respectively. We assessed associations between EAH and BMI z-score using Pearson correlations. We included 196 participants (50% girls; mean (SD) 11.9 (2.3) years old) for the first completion and 153 for the second completion. The exploratory factor analysis generated the same three subscales as the original questionnaire: negative affect (α = 0.86; ICC = 0.78), fatigue/boredom (α = 0.75; ICC = 0.70), and external cues (α = 0.68; ICC = 0.54). Participant's BMI z-scores were positively associated with the average scores from the negative affect subscale (r = 0.19; ρ = 0.009). Our results suggest that this questionnaire has an adequate construct validity, internal consistency, and temporal stability.

Invasive group A streptococcal infection outbreaks of typeemm118 in a long-term care facility, and of type emm74 in the homeless population, Montréal, Quebec.

BACKGROUND: Two invasive group A streptococcus (iGAS) infection outbreaks occurred in Montreal in 2016 and 2017; one in a long-term care facility (typeemm118) and one in the community, primarily involving homeless people (typeemm74). OBJECTIVE: To describe two recent iGAS outbreaks in Montréal and highlight the challenges in dealing with these outbreaks and the need to tailor the public health response to control them. METHODOLOGY: All cases of iGAS were investigated and the isolates were sent to the laboratory foremmtyping. In both outbreaks, cases of superficial group Astreptococcus(GAS) infection were identified, through 1) systematic case detection accompanied by screening for asymptomatic carriers among residents and employees of the long-term care facility and 2) sentinel surveillance among homeless people. Visits were made to community organizations providing homeless services (including shelters) and social networks were analyzed to establish whether there were any links among cases of GAS infection (both invasive and noninvasive) and locations frequented. In both outbreaks, recommendations were made to service providers regarding enhancement of infection prevention and control measures. RESULTS: In the long-term care facility, five cases of typeemm118 iGAS were identified over a 22-month period, one of which resulted in death. All residents were screened and no carriers were identified. Among the employees, 81 (65%) were screened and fourcarriers were identified. Of those, one was a carrier of typeemm118 GAS. All carriers were treated, and subsequent follow-up sampling on three carriers (including the one withemm118) was negative.In the community, 23 cases of typeemm74 iGAS were detected over a 16-month period, four of which resulted in death. Half of the cases (n=12) were described as homeless, and six others were users of services for the homeless. Sentinel surveillance of superficial infections yielded 64 cultures with GAS, chiefly on the skin, including 51 (80%) of typeemm74. An analysis of the social networks revealed the large number and variety of resources for the homeless used by the cases. Visits to the community organizations providing homeless services revealed the heterogeneity and precariousness of some of these services, the difficulties encountered in applying adequate health and hygiene measures, and the high degree of mobility amongst those who use these services. CONCLUSION: The detection and control of iGAS outbreaks in both long-term care establishments and among community organizations providing homeless services are very complex. An outbreak of iGAS can develop in the background over a long time and be easily overlooked despite cases being admitted to the hospital.Emmtyping and systematic research of previous cases of iGAS are essential tools for the detection and characterization of outbreaks. Close cooperation among public health agencies, clinical teams, community organizations and laboratories is essential for proper monitoring and the reduction of GAS transmission in the community and health care settings.

Bispecific monoclonal antibodies produced by somatic cell fusion increase the potency of tissue plasminogen activator.

Bispecific monoclonal antibodies that bind simultaneously to human fibrin and tissue plasminogen activator (tPA) enhance the fibrinolytic potency of tPA. Two bispecific antibodies (F36.23 and F32.1) were generated by somatic cell fusion. Antibody F36.23 derives its tPA binding from monoclonal anti-tPA antibody TCL8 and its fibrin binding from monoclonal antifibrin antibody 59D8. After purification from cell supernatants and ascites by two steps of affinity chromatography, hybrid-hybridoma bispecific antibody F36.23 simultaneously bound tPA and fibrin in solution and in solid-phase assays. In an assay for the lysis of human fibrin monomer, F36.23 increased the fibrinolytic potency of tPA by 5 to 10 fold, regardless of whether the bispecific antibody had been combined with the tPA before or during the assay. Bispecific F36.23 F(ab')2 also bound tPA and fibrin simultaneously, and the enhancement in fibrinolysis in the presence of F36.23 F(ab')2 was identical to that in the presence of intact F36.23. The second bispecific antibody, F32.1, was produced by an alternative strategy that has a wider potential for application in other systems. Hybridoma bispecific antibody F32.1 was derived from the fusion of immune splenocytes (in mice immunized with a synthetic oligopeptide representing the amino terminus of the alpha-chain of human fibrin) with the anti-tPA cell line TCL8. The properties of hybridoma bispecific antibody F32.1 and its F(ab')2 were indistinguishable from those of hybrid-hybridoma bispecific antibody F36.23 in solid-phase binding assays and in assays of fibrinolysis. Bispecific antibodies produced by somatic cell fusion, particularly in the form of F(ab')2, may have potential for use in clinical thrombolysis.

The effect of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on cultured dog pancreatic duct epithelial cells.

To study the morphologic and genetic events associated with the carcinogenic process in the pancreas, we have isolated and cultured a cell line of dog pancreatic duct epithelial cells and treated these cells with a carcinogen. The pancreatic duct epithelial cells were plated onto Vitrogen-coated Transwell inserts suspended above a feeder layer of human gallbladder myofibroblasts. The epithelial cells grew steadily into polarized monolayers, could be passaged repeatedly, and demonstrated the typical morphologic, immunohistochemical, and flow cytometric profile of normal well-differentiated columnar pancreatic epithelial cells. After being treated with 10(-5) M N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) for 48 h, the treated cells grew on plastic surfaces. When grown in organotypic culture, the MNNG-treated cells were cuboidal with a multilayered, pseudostratified architecture. Flow cytometry demonstrated aneuploidy and a high percentage of the cells in S phase after reaching confluency, in sharp contrast to untreated cells. Cytogenetic analysis of the MNNG-treated cells revealed frequent chromosomal trisomy and tetrasomy. The secretion of mucin was also different in the MNNG-treated cells versus the untreated cells. The cultured pancreatic epithelial cells may be useful as an assay system to study the genotoxicity of known and potential carcinogens.

Growth and function of isolated canine pancreatic ductal cells.

These studies investigated the growth characteristics and functional properties of isolated canine pancreatic ductal epithelial cells. Cells were isolated from the accessory pancreatic duct and cultured by using three conditions: on vitrogen-coated petri dishes with fibroblast conditioned medium (nonpolarized); in vitrogen-coated Transwells above a fibroblast feeder layer (polarized); or as organotypic rafts above a fibroblast-embedded collagen layer (polarized). Growth characteristics, transepithelial resistances, and carbonic anhydrase and cyclic adenosine monophosphate (AMP) responses were evaluated. Under polarized conditions, the cells grew as monolayers with columnar epithelial characteristics. The monolayers developed high transepithelial resistance and became impervious to the passage of horseradish peroxidase. Epithelial growth factor (EGF) (2 ng/ml) stimulated ductal cell growth and accelerated the formation of a high-resistance monolayer. Forskolin (10 microM) rapidly decreased transepithelial resistance. Carbonic anhydrase activity, which was lower in nonpolarized compared with polarized conditions, was stimulated by carbachol (175 microM). Secretin, however, did not stimulate carbonic anhydrase activity in these cells. Although secretin stimulated adenylyl cyclase activity in early-passage cells, this response was lost in later-passage cells. Both vasoactive intestinal polypeptide (VIP; 1 microM) and forskolin (10 microM) consistently increased adenylyl cyclase activity. Isolated canine pancreatic ductal epithelial cells proliferate in vitro, develop high-resistance epithelial monolayers, and respond to stimuli that activate adenylyl cyclase. These cells should provide a useful model for regulatory studies of ductal cell functions.

High-level expression of antibody-plasminogen activator fusion proteins in hybridoma cells.

We show that the mouse gamma 2b heavy chain or human beta-globin 3' untranslated region can greatly enhance protein expression in myeloma cells transfected by genes coding for antibody-plasminogen activator fusion proteins. Expression plasmids were constructed containing a cloned genomic heavy chain variable region from fibrin-specific monoclonal antibody 59D8, a cloned genomic constant region of the mouse gamma 2b heavy chain, and DNA sequence coding for either tissue-type plasminogen activator (tPA) or a segment of urokinase (UK) and their respective 3' untranslated sequences. Cell lines transfected with these constructs, pSVtPA (tPA) and pSVUKG(UK), produced extremely low levels of mRNA and protein (0.008-0.06 micrograms/ml) in comparison with the parental 59D8 myeloma cell line (7.6-10 micrograms/ml). In vitro nuclear run-off analysis indicated that the low steady-state levels of mRNA encoded by pSVUKG(UK) did not result from a lower rate of transcription of the transfected gene (relative to the rate of transcription of the endogenous heavy chain gene in the 59D8 parent cells). In an attempt to increase protein secretion, we assembled the expression plasmids pSVtPA(Ig), pSVUKG(Ig), and pSVUKG(beta), in which the 3' untranslated region of the mouse gamma 2b heavy chain or human beta-globin gene was substituted for the 3' untranslated region of the plasminogen activator gene. Analysis of supernatant media from cell lines transfected with these constructs showed an increase in recombinant protein secretion of 68 to 100 fold in comparison with that from cell lines transfected with pSVtPA(tPA) or pSVUKG(UK).

Reconstituted human oral and esophageal mucosa in culture.

We have successfully established monolayer and organotypic culture techniques for growing human oral and esophageal epithelial cells. Cells in monolayer culture were grown in serum-free medium, modified from techniques previously reported by our group. The organotypic cultures were grown in a defined medium supplemented with 10% fetal calf serum. Oral and esophageal cells were maintained in keratinocyte basal medium with pituitary extract and other supplements, and 0.05 mM calcium for 7-9 and 9-11 passages, respectively. Both cell types had similar morphology by phase contrast microscopy. When confluent, the cells were predominantly small, basaloid, and uniform and interspersed with larger, differentiated cells. By immunohistochemistry, both cell types in monolayer were positive to AE1, AE3, and 34BE12 antibodies to keratins of stratified epithelia. Oral epithelial cells in monolayer also were positive to 35BH11, representative of simple epithelial keratins, while esophageal cells were not. The esophageal cells were focally positive to K13, while the oral cells were negative. Both were negative for K19. When comparing monolayer to organotypic cultures and to in vivo specimens, there was a significant difference in the expression of keratins. Using organotypic cultures, AE1, AE3, and 34BE12 were strongly positive in both oral and esophageal cells, similar to in vivo tissues. In contrast to monolayers, both were also focally positive for K19. Esophageal cells were strongly positive for K13, while the oral cells were mildly but uniformly positive. Both were negative for keratins of simple epithelia. These two cell culture techniques offer unique opportunities to study the pathobiology, including carcinogenesis, of stable cell systems from the oral and esophageal epithelia.

Culture of human main pancreatic duct epithelial cells.

Attempts to grow human pancreatic duct epithelial cells in long-term culture have proven difficult. We have developed a system of growing these cells for several passages by adapting methods used to culture dog pancreatic duct cells. Epithelial cells were enzymatically dissociated from the main pancreatic duct and plated onto collagen-coated culture inserts suspended above a human fibroblast feeder layer. After primary culture, the cells were either passaged onto new inserts or plastic tissue culture plates in the absence of collagen. Cells grown on the latter plates were maintained in a serum-free medium. Primary pancreatic duct epithelial cells grow steadily to confluence as a monolayer in the feeder layer system. After primary culture, cells passaged onto new inserts with fresh feeder layer or plastic plates and fed with serum-free medium continued to develop into confluent monolayers for up to four passages. The cells were columnar with prominent apical microvilli, sub-apical secretory vesicles, and lateral intercellular junctions resembling the morphology of normal in vivo epithelial cells. These cells were also positive for cytokeratin 19, 7, and 8 and carbonic anhydrase II, as measured by immunohistochemistry. Metabolically, these cells synthesized and secreted mucin, as measured by incorporation of tritiated N-acetyl-D-glucosamine. In conclusion, we demonstrated that human pancreatic epithelial cells from the main duct can be successfully grown in culture and repeatedly passaged using a feeder layer system, with serum-free medium, and in organotypic cultures.

Isolation and long-term culture of gallbladder epithelial cells from wild-type and CF mice.

Mice with targeted disruption of the cftr gene show pathophysiologic changes in the gallbladder, which correlate with hepatobiliary disease seen in cystic fibrosis patients. As gallbladder epithelium secretes mucin, and as this epithelium consists of a relatively homogenous cell type, study of CFTR function in these cells would be beneficial to delineate the complex cellular functions of this protein. The size and anatomic location of the murine gallbladder makes such studies difficult in vivo. Therefore, the need exists for in vitro models of gallbladder epithelium. We describe a method to isolate and culture murine gallbladder epithelium from wild-type and CF mice. Cells were grown in a monolayer on porous inserts over a feeder layer of fibroblasts. These nontransformed cells can be successively passaged and maintain a well-differentiated epithelial cell phenotype as shown by morphologic criteria, characterized by polarized columnar epithelial cells with prominent microvilli and intercellular junctions. Organotypic cultures showed columnar cells simulating in vivo morphology. This culture system should be valuable in delineating cellular processes relating to CFTR in gallbladder epithelium.

Chiral identification and determination of ephedrine, pseudoephedrine, methamphetamine and methcathinone by gas chromatography and nuclear magnetic resonance.

The enantiomers of the related substances methamphetamine, ephedrine, pseudoephedrine and methcathinone were determined by both gas chromatography after derivatization and by nuclear magnetic resonance using a chiral solvating agent. For GC the substances were derivatized with (R)-(+)-alpha-methoxy-alpha-(trifluoromethyl)phenylacetic acid (MTPA) to give diasteromeric derivatives. Resolution (baseline) of at least 1.6 was obtained between all derivatives. NMR determination of the enantiomers was conducted in a chiral environment by the addition of the chiral solvating agent, (R)-(+)-1,1'-bi-2-naphthol, to NMR solutions of the substances. Racemization of methcathinone was demonstrated to be facile by exposure to alkaline solutions for varying periods of time. Enantiomeric ratios of some products derived from the oxidation of ephedrine were determined.

Cefprozil versus Cefuroxime Axetil in the Treatment of Acute Sinusitis.

The objective of this multicentre, randomised, open-label, general practice (GP) study was to evaluate the efficacy and tolerability of cefprozil (Cefzil(trade mark), Bristol-Myers Squibb) compared with that of cefuroxime axetil (Ceftin((R)), Glaxo Wellcome) in the treatment of adult subjects with acute sinusitis. Typical of the GP setting, diagnosis was made based solely on clinical signs and symptoms of acute disease. Sinus radiography was performed post-randomisation. A total of 381 adolescent and adult patients were randomly assigned to 10 days' treatment with either cefprozil, 500mg orally twice daily (n = 191), or cefuroxime axetil, 250mg orally twice daily (n = 190). Based on predefined criteria, treatments were found to be equally effective in terms of proportions of patients in the per-protocol population that were cured, improved or failed (p = 0.20). Similar results were observed when the evaluation was performed on the subset of patients with radiographic evidence of sinusitis and when the evaluation was based on the investigator's judgement. Similar rates of adverse events were observed in the two treatment groups. In summary, cefprozil 500mg twice daily is as well tolerated and as effective as cefuroxime axetil 250mg twice daily for the treatment of adolescent and adult patients with clinical signs and symptoms of acute sinusitis.

Effect of moderate intake of sweeteners on metabolic health in the rat.

The rise in prevalence of obesity, diabetes, metabolic syndrome, and fatty liver disease has been linked to increased consumption of fructose-containing foods or beverages. Our aim was to compare the effects of moderate consumption of fructose-containing and non-caloric sweetened beverages on feeding behavior, metabolic and serum lipid profiles, and hepatic histology and serum liver enzymes, in rats. Behavioral tests determined preferred (12.5-15%) concentrations of solutions of agave, fructose, high fructose corn syrup (HFCS), a combination of HFCS and Hoodia (a putative appetite suppressant), or the non-caloric sweetener Stevia (n=5/gp). HFCS intake was highest, in preference and self-administration tests. Groups (n=10/gp) were then assigned to one of the sweetened beverages or water as the sole source of liquid at night (3 nights/wk, 10wks). Although within the normal range, serum cholesterol was higher in the fructose and HFCS groups, and serum triglycerides were higher in the Agave, HFCS, and HFCS/Hoodia groups (vs. water-controls, p<0.05). Liver histology was normal in all groups with no evidence of steatosis, inflammation, or fibrosis; however serum alanine aminotransferase was higher in the fructose and HFCS groups (vs. water-controls, p<0.05). Serum inflammatory marker levels were comparable among Stevia, agave, fructose, HFCS, and water-consuming groups, however levels of IL-6 were significantly lower in association with the ingestion of Hoodia. There were no differences in terminal body weights, or glucose tolerance assessed by 120-min IVGTTs performed at the end of the 10-week regimen. We conclude that even moderate consumption of fructose-containing liquids may lead to the onset of unfavorable changes in the plasma lipid profile and one marker of liver health, independent of significant effects of sweetener consumption on body weight.

Identification of N-formylnorcocaine and N-benzoylnormethylecgonine in illicit cocaine.

Gas chromatographic determination of the acidic and neutral components of illicit cocaine indicated the presence of one or more common components in different samples. Isolation and examination of the spectroscopic properties of the major impurity indicated it to be N-formylnorcocaine. The material was compared with authentic material synthesized from norcocaine. N-Benzoylnormethylecgonine was also found to be present in illicit cocaine.

PGE generates intracellular cAMP and accelerates mucin secretion by cultured dog gallbladder epithelial cells.

Mucin is the main secretory product of gallbladder epithelial cells. Increased gallbladder mucus secretion has been implicated in gallstone formation in humans. The mechanisms underlying control of mucin synthesis and secretion by the gallbladder are not known. This study aimed to elucidate the efficacy of a panel of secretagogues to stimulate mucin secretion and to determine the intracellular second messengers involved. Studies were carried out on normal well-differentiated epithelial cells from dog gallbladder grown in monolayer culture. Intracellular adenosine 3',5'-cyclic monophosphate (cAMP) as measured by radioimmunoassay increased in response to prostaglandin (PG) E2, PGE1, vasoactive intestinal peptide, epinephrine, and isoproterenol. The greatest effect, a 37-fold increase in cAMP level, was noted with PGE2 at 1.0 microM concentration. In contrast, three breakdown products of phosphatidylinositol (inositol triphosphate, inositol bisphosphate and inositol monophosphate) were not detected with any of the secretagogues tested. Assay of mucin secretion using tritiated N-acetyl-D-glucosamine, a mucin precursor, showed that the same secretagogues noted to increase intracellular cAMP led to an increase in mucin secretion. No correlation was noted, however, between the magnitude of the intracellular cAMP rise and the amount of mucin secreted. A membrane-permeable form of cAMP, dibutyryl cAMP, mimicked PGE2-induced mucin secretion. The results unequivocally show that secretagogue-stimulated mucin secretion in these normal gallbladder epithelial cells can proceed via a cAMP signal transduction pathway.

Stability of methadone in four vehicles for oral administration.

The stability of methadone in vehicles commonly used for oral administration was determined. Solutions of methadone were prepared in (1) orange-flavored Tang, (2) grape-flavored Kool-Aid, (3) apple juice, (4) grape-flavored Crystal Light, and (5) grape-flavored Crystal Light plus 0.1% sodium benzoate. For each of the first four preparations listed, two solutions were formulated at each methadone hydrochloride concentration of 0.2, 0.8, and 1.5 mg/mL; one set of three concentrations of each solution was stored in a refrigerator at 5 degrees C for up to 55 days, and the other set was stored unprotected from light at 20-25 degrees C for up to 17 days. Only three Crystal Light plus sodium benzoate solutions were prepared at the same methadone concentrations and stored at 20-25 degrees C for up to 29 days. Methadone concentrations were measured by high-performance liquid chromatography. Methadone was stable (loss of potency, less than 5%) for up to 17, 11, 9, and 8 days when stored at 20-25 degrees C in Kool-Aid, Tang, apple juice, and Crystal Light, respectively, and for up to 29 days when stored at 20-25 degrees C in Crystal Light plus sodium benzoate. Methadone was stable for up to 55, 49, 47, and 34 days when stored at 5 degrees C in Kool-Aid, Tang, apple juice, and Crystal Light, respectively. All the solutions that did not contain sodium benzoate and were stored at room temperature displayed unacceptable microbial growth after approximately two weeks. No significant loss of methadone potency occurred in any of the vehicles for oral administration during the study.

Presence of conjugated catecholamines in rat brain: a new method of analysis of catecholamine sulfates.

A new method of measuring catecholamine (CA) sulfate permitted us to detect its presence in rat brain for the first time. The procedure consisted of separating the CA sulfate from the free CA by alumina adsorption followed by passage through Dowex, and measuring the CA sulfate by a radioenzymatic assay in the presence of a sulfatase. This method permitted demonstration of the presence of dopamine sulfate, and occasionally, of norepinephrine and epinephrine sulfate in the hypothalamus, striatum, and hippocampus of rat brain.