Adaptations of an Effective Evidence-Based Pediatric Weight Management Intervention.

Current childhood obesity treatment programs do not address medically underserved populations or settings where all members of an interdisciplinary team may not exist-either within one organization or within the community. In this paper, we describe the use of a community-academic partnership to iteratively adapt Epstein's Traffic Light Diet (TLD), into Building Healthy Families (BHF), a community-placed evidence-based pediatric weight management intervention (PWMI) and evaluate its effectiveness in reducing BMI z scores. Nine cohorts of families completed BHF. Participants included children aged 6-12 years with obesity (M = 9.46, SD = 1.74). The Framework for Reporting Adaptations and Modifications-Expanded guided our classification of modifications across BHF cohorts. Using the FRAME reporting structure, the changes that were documented were (1) planned and occurred pre-implementation, (2) based on decisions from local stakeholders (e.g., school administrator, members of the implementation team), and (3) specific to changes in content and context-with a focus on implementation and potential for local scale-up. The nature of the adaptations included adding elements (whole of family approach), removing elements (calorie counting), and substituting elements (steps for minutes of physical activity). Across 9 cohorts, 84 families initiated the BHF program, 69 families successfully completed the 12-week program, and 45 families returned for 6-month follow-up assessments. Results indicated that the BMI z score in children was reduced by 0.31 ± 0.17 at 6 months across all cohorts. Reduction in BMI z score ranged from 0.41 in cohort 4 to 0.13 in cohort 5. Iterative adaptations to BHF were completed to improve the fit of BHF to the setting and participants and have contributed to a sustained community PWMI that adheres to the underlying principles and core elements of other evidence-based PWMIs. Monitoring adaptations and related changes to outcomes can play a role in long-term sustainability and effectiveness.

Molecular Analysis of Diepoxybutane-Induced Mutations at the rosy Locus of Drosophila melanogaster.

We have analyzed at the molecular level diepoxybutane-induced mutants determined to have lesions affecting expression of the ry locus. Of the 21 mutants analyzed here, genetic analysis suggested that five were putative deficiencies involving ry and adjacent lethal loci. However, molecular analysis confirmed that only two of these five putative deficiencies were in fact deletions detectable by the methods used in the analysis. The remaining 16 mutants were viable as homozygotes, suggesting that their lesions were confined to the ry locus. Seven of these 16 intragenic mutants were determined to be deletions of genetic material as evidenced by altered restriction patterns relative to the wild type patterns. Thus, nine of 21 (43%) diepoxybutane-induced mutants are due to deletions ranging in size from approximately 50 base pairs to more than 8 kilobase pairs. Most of the deletions (seven of nine or 78%) are intragenic and less than 250 base pairs in size; it seems that most, if not all, affect coding rather than regulatory sequences.

Physical organization of the metJB component of the Escherichia coli K-12 metJBLF gene cluster.

The structures of a series of plaque-forming metJB transducing phage were studied by restriction endonuclease mapping and enzyme activity assay. One of these phage, lambda pmet100, was inactivated by heat shock in the presence of EDTA, and deletion mutants were selected from the survivors. Two of these mutants, lambda pmet100 delta 1 and lambda pmet100 delta 2, were used to confirm the gene order metJ metB when moving clockwise on the linkage map of Escherichia coli K-12. Additional results indicate that the metB gene can be expressed independently of any other component of the met regulon and that the metJ gene also forms a separate transcription unit.

Typing of herpes simplex virus with type-specific human immunoglobulin M in an indirect immunofluorescence assay.

Sera from nine individuals with suspected primary herpes simplex virus type 1 (HSV-1) or type 2 (HSV-2) infection were screened to identify those containing HSV type-specific immunoglobulin M (IgM). Selected sera were then utilized in an IgM-specific indirect immunofluorescent-antibody HSV-typing assay (patent pending). To evaluate the procedure, 29 HSV isolates were grown in cultures of continuous human amnion cells, fixed, and used as substrates for indirect immunofluorescence. Determination of virus type was based on intensity of fluorescence of the substrate with HSV-1- and HSV-2-specific antisera after staining with fluorescein-conjugated anti-human IgM. Typing of the isolates by restriction endonuclease digestion showed that of 29, 18 were HSV-2 and 11 were HSV-1. Results by IgM-specific indirect immunofluorescent-antibody assay were identical to those by restriction endonuclease digestion for 27 of the isolates; 2 isolates failed to replicate adequately in the test cells. The IgM-specific indirect immunofluorescent-antibody procedure appears to be a simple, rapid, and accurate technique which could be of use to clinical virology laboratories.

Effect of viral infections on pulmonary function in patients with chronic obstructive pulmonary diseases.

The effect of 100 separate viral infections of the respiratory tract on pulmonary function was evaluated prospectively over an eight-year period in 84 patients with chronic obstructive pulmonary diseases and in eight normal subjects. Some viral infections were associated with small acute declines in forced vital capacity and/or 1-sec forced expiratory volume of 25-300 ml. These declines were detectable only during the 90-day period after infection. The greatest abnormalities of pulmonary function followed infections with influenza virus, and the mean acute changes in 1-sec forced expiratory volume (-118.5 ml) were significantly greater than expected (-15.2 ml; P = 0.03). Smaller, statistically insignificant declines followed infections with parainfluenza virus, rhinovirus, adenovirus, and respiratory syncytial virus, and no changes were detectable after infections with coronavirus, herpes simplex virus, Mycoplasma pneumoniae, and Haemophilus influenzae. Long-term effects of influenza or other viral infections on the course of chronic obstructive pulmonary disease were not detected in this study population.

Association of viral and Mycoplasma pneumoniae infections with acute respiratory illness in patients with chronic obstructive pulmonary diseases.

The relative importance of respiratory virus and M. pneumoniae infections as causes of acute respiratory illnesses was studied over an 8 yr period in 150 subjects who were normal or who had varying degrees of chronic obstructive pulmonary disease (COPD). Viral or M. pneumoniae infections were associated with 186 of 1,030 (18%) illnesses studied, whereas these infections were detected in only 86 of 1,398 (6%) illness-free periods (P less than 0.01). Rhinoviruses, influenza viruses, parainfluenza viruses and coronaviruses were each significantly associated with acute respiratory illnesses. The occurrence of acute respiratory illnesses and viral infections was the same in subjects with moderate to severe COPD as it was in subjects who were normal or who had mild disease.

Variables associated with changes in spirometry in patients with obstructive lung diseases.

One hundred fifty subjects were enrolled in a long-term study of obstructive lung diseases; 84 of these were subjected to five or more spirometric studies over a period of two or more years. Stepdown regression analysis was performed to determine the association between many different variables and the annual rates of change in the forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1). The following associations were noted to be significant (p less than 0.03); more favorable rates of change of the FVC and FEV1 were associated with a higher alpha1-antitrypsin level and older age. Less favorable changes were associated with more years of cigarette smoking, more airway reactivity and more frequent lower respiratory tract illnesses.

Haemophilus influenzae and haemophilus parainfluenzae in chronic obstructive pulmonary disease.

The importance of nonencapsulated strains of Haemophilus influenzae and H. parainfluenzae in the pathogenesis of chronic obstructive pulmonary disease (C.O.P.D.) has been investigated in 150 patients observed at two-month intervals from 1968 to 1975. H. influenzae was distinguished from H. parainfluenzae by demonstrating requirements for both X and V factors. H. influenzae was isolated more often from sputum cultures from patients with severe C.O.P.D. (13.8%) than from patients with mild disease (4.4%, P less than 0.01). In contrast, H. parainfluenzae was isolated with equal frequency from sputums of patients with mild (40%) or severe (43%) disease. H. influenzae was present in the sputum more often during exacerbations of acute respiratory illness (15.4%) than during sympton-free periods (9.6%, P less than 0.01), while isolation rates of H. parainfluenzae did not increase during periods of illness (35% versus 39%). Antigens specific for the H. influenzae species and for the H. parainfluenzae species were used in a complement-fixation test to detect antibody rises in sera collected from the patient population. Fourfold or greater rises in titre of antibodies to H. influenzae were detected on 76 occasions in 53 patients, and 46 of these rises were associated with acute respiratory illnesses. In contrast, no antibody rises were detected with the H. parainfluenzae antigen. These observations indicate that nonencapsulated strains of H. influenzae may have pathogenic potential in patients with C.O.P.D., whilst H. parainfluenzae should be considered as normal flora.

Immunological nature of antimycobacterial phenomenon in macrophages.

Heptane-extractable fractions (HEF) prepared from immune-activated macrophages (IA-M) of tubercle bacilli or bovine gamma globulin-sensitized and -challenged guinea pigs inhibited the growth of tubercle bacilli whereas HEF of normal macrophages exerted no antibacterial activity. In distinction to the strong antibacterial activity of HEF of IA-M, HEF of immune macrophages exerted weak or no antimycobacterial activity. HEF of alveolar macrophages exerted stronger antibacterial activity than HEF of peritoneal macrophages. The degree of the antibacterial activity of HEF was determined primarily by the time of macrophage collection from antigenically stimulated animals. The antibacterial activity gradually increased and peaked at 2 weeks after the antigenic stimulation of sensitized animals; subsequently, the activity declined and disappeared in about 5 weeks. Similar to other immunological reactions, the stimulation of sensitive animals with specific antigen induced an anamnestic reaction which was characterized by a rapid recall of the macrophage antimycobacterial phenomenon (MAP). The antibacterial strength of the recalled phenomenon in sensitized animals was dependent upon the intensity of the sensitizing regimen; the phenomenon was much stronger in three times-sensitized animals than in once- and twice-sensitized animals. The time of appearance and the specificity of induction and of recall of the MAP indicate that the phenomenon is associated with the activated state in macrophages and, as a consequence of this association, it has a well-defined immunological nature.

Role of mycobactin in the growth and virulence of tubercle bacilli.

Tubercle bacilli failed to grow in iron-void media enriched with solutions of iron-containing transferrin (Tr) or ferritin (F) because these substances do not provide the bacilli with iron, which is essential for their growth. Animal serum and macrophages possessed no iron carrier with an ability to satisfy the need of the bacteria for the metal. Mycobactin (M), the growth-product of tubercle bacilli, removed iron from Tr and F and supplied the metal for bacillary utilization. The role of M in the growth of tubercle bacilli was influenced by nonionic surfactants which inhibited bacillary growth by removing M from the bacillary cells and interfering with the absorption of M-iron complexes. Experiments with Tween 80, Triton WR-1339, and lecithin showed that avirulent bacilli lose M at lower concentrations of the surfactants than virulent bacilli. Since avirulent and virulent bacilli possess the same amount of M, these findings indicate that M is bound more firmly to lipid-rich virulent than lipid-poor avirulent cells. These findings indicate that the resistance of virulent bacilli to the M-removing activity of the surfactants is an indicator of their ability to multiply in the infected host and may be used as a measure of bacillary virulence.

Antimycobacterial effect of lysates prepared from immunologically activated macrophages.

Lysates or heptane extracts of peritoneal (P) and alveolar (A) normal macrophages (N-M), immune macrophages (I-M), and immune-activated macrophages (IA-M) were examined for antimycobacterial activity by the agar-plate diffusion test. This test has been found suitable to reveal the antibacterial activity in 3-day incubated, but not in freshly prepared, lysates. Results showed that materials of IA-AM or I-AM and of IA-PM exerted antimycobacterial effects, whereas materials of N-PM, I-PM, and of N-AM were usually inactive. Antimycobacterial activity of lysates of AM was stronger than that of PM. The formation of antibacterial factors during an incubation of M lysates, the solubility of the factors in heptane, and various other characteristics suggested that the antimycobacterial effect was caused by the formation of toxic levels of non-esterified fatty acids. M lysates exerted equal activities against BCG, H(37)Ra, and H(37)Rv strains of tubercle bacilli. The presence of antimycobacterial activity in lysates prepared from IA-M of either BCG- or BCG-sensitized animals indicated that the potential to generate antimycobacterial activity is associated with the state of delayed hypersensitivity and the state of activation of M.

Employment of tuberculostasis in serum-agar medium for the study of production and activity of Mycobactin.

Mycobactin (M), an iron-chelating product of tubercle bacilli, neutralized serum tuberculostasis by removing growth-essential iron from transferrin (Tr) and supplying the metal to the bacteria. The competition for iron between Tr and M has been demonstrated by the agar-plate diffusion test. This test is suitable not only for the study of Tr-iron-M interplay but also for the evaluation of serum tuberculostasis. Extremely poor solubility of M in water and consequently its association with lipoidal cell wall of tubercle bacillus was overcome by the use of water-dispersible and surface-active Tween 80. The addition of Tween 80 to culture media insured the presence of M in spent media; otherwise M was extracted from bacillary cells with a solution of Tween 80 or a mixture of ethanol and Tween 80. Although M was produced irrespective of the amount of iron present in culture medium, its production in iron-poor medium was more prolific than in iron-rich medium. M extracted from BCG or H(37)Rv cells neutralized serum tuberculostasis as effectively for the homologous as for heterologous strains. However, the extract of virulent bacilli was much more active in the neutralization than similar extract prepared from attenuated cells; whether this difference is of quantitative or qualitative nature remains to be determined.

Mechanism of Tuberculostasis in Mammalian Serum III. Neutralization of Serum Tuberculostasis by Mycobactin.

Tubercle bacilli exposed to an iron-poor medium multiplied at a slower rate but released more of the serum-tuberculostasis neutralizing factor (TNF) than the bacilli in an iron-rich medium. This growth-promoting factor found in spent medium exhibited characteristics which suggested its relationship to or identity with mycobactin. The identity of these two bacillary products was established by showing that both iron-free mycobactin and TNF promoted bacillary multiplication in tuberculostatic serum. This study resolved a long-standing controversy as to whether mycobactin serves as a growth factor or as a carrier of iron for tubercle bacilli. It was found that the tuberculostasis in mycobactin-neutralized serum was reconstituted by the addition of iron-free transferrin (Tr). The investigation of the interplay between mycobactin and Tr revealed that mycobactin does not serve as a growth factor but as a carrier of growth-essential iron which mycobactin (as contrasted to Tr) provides to tubercle bacilli in a utilizable form.

Mechanism of tuberculostasis in mammalian serum. II. Induction of serum tuberculostasis in guinea pigs.

The growth of tubercle bacilli in serum samples of untreated animals depends upon the availability of ionic iron which serves as a growth factor in supporting bacillary multiplication. The amount of available iron in serum is determined by the ratio between iron-saturated and iron-free transferrin; a low value for the ratio is associated with tuberculostasis (e.g., human serum, 0.4), whereas a high value is associated with the growth-supporting quality (e.g., guinea pig serum, 5.6). The treatment of guinea pigs with lipopolysaccharide of Escherichia coli or tuberculous cell wall material consistently and significantly reduced serum iron levels; a similar but less striking effect was observed in BCG-vaccinated animals. Pronounced differences were observed in the time of appearance and duration of serum hypoferremia; in lipopolysaccharide-treated animals, it appeared in 1 day and lasted for several days, whereas in BCG-vaccinated animals it appeared in about 2 weeks and lasted for much longer time periods. The induced hypoferremia was always associated with the concomitant development of serum tuberculostasis which could be neutralized by the addition of iron. These results indicate, therefore, that the mechanism of induced serum tuberculostasis in lipopolysaccharide- or tuberculous cell wall-treated and BCG-vaccinated guinea pigs is the same as that present in tuberculostatic sera of untreated animals.