ISCEV and IPS guideline for the full-field stimulus test (FST).

The full-field stimulus test (FST) is a psychophysical technique designed for the measurement of visual function in low vision. The method involves the use of a ganzfeld stimulator, as used in routine full-field electroretinography, to deliver full-field flashes of light. This guideline was developed jointly by the International Society for Clinical Electrophysiology of Vision (ISCEV) and Imaging and Perimetry Society (IPS) in order to provide technical information, promote consistency of testing and reporting, and encourage convergence of methods for FST. It is intended to aid practitioners and guide the formulation of FST protocols, with a view to future standardisation.

Huge Intraosseous Tibial Haemangioma Managed with Embolisation, Excision and Fibular Ilizarov Reconstruction: A Case Report.

AIM: Our aim is to report the successful treatment of an intraosseous haemangioma of tibia with an atypical presentation through a multidisciplinary approach of preoperative embolisation, a subtotal resection of the tibia and subsequent reconstruction with the Ilizarov medial fibular translation technique. BACKGROUND: En bloc excision is the treatment of choice for large tumours of the tibia. However, there is no single recommended method for the reconstruction of the resulting bony defect. CASE: A 22-year-old female presented with a massive intraosseous haemangioma of the entire tibia. Sequential, multimodal treatment consisted of (1) preembolisation, (2) en bloc resection and (3) reconstruction of the extensive skeletal defect via the Ilizarov method of fibular medialisation. Radiologic union occurred at 6 months and graft hypertrophy at 22 months. At 45 months, the patient was fully weight-bearing without need for an assistive device. CONCLUSION: Resection and reconstruction of a large intraosseous haemangioma of the tibia can be treated successfully using a well-planned sequential management of embolisation, resection and Ilizarov fibular grafting. SIGNIFICANCE: This report highlights the successful management of an unusually extensive and difficult tumour through appropriate and meticulous perioperative multidisciplinary planning, execution and follow-up. HOW TO CITE THIS ARTICLE: Barsales KAD, Javier J, Catibog JJ, et al. Huge Intraosseous Tibial Haemangioma Managed with Embolisation, Excision and Fibular Ilizarov Reconstruction: A Case Report. Strategies Trauma Limb Reconstr 2021;16(1):60-63.

Broad-spectrum Antimicrobial Activity of Purified Hemocyanin Subunit IIIA Isolated from Asian Horseshoe Crab, Tachypleus gigas.

BACKGROUND AND OBJECTIVES: Hemocyanin Subunit IIIA is believed to possess antimicrobial properties, but its efficacy against microbial pathogens is still unclarified. Thus, this study aimed to determine antimicrobial activities of hemocyanin subunit IIIA and to identify the best activator of this protein. MATERIALS AND METHODS: The hemocyanin was partially purified using spin column affinity, its fraction was applied to Hi-Prep Sephacryl Exclusion 26/60 2-200 HR column, followed by Hi-Prep 26/10 Desalting Column on fast protein liquid chromatography. The purity of hemocyanin was validated by Matrix Assisted Laser Desorption Ionization-Time of Flight/Mass Spectrometry. The antimicrobial activity was performed by Disc Diffusion Test. RESULTS: Purified hemocyanin subunit IIIA was identified to have a molecular weight of 72.9 kDa. SDS was found to be the best activator of hemocyanin, as indicated by elevated level of phenoloxidase. As for antimicrobial activity, hemocyanin was minimally inhibited by all bacteria strains tested (Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae), with relatively lower Minimum Inhibitory Concentration (MIC) at 0.005 g mL-1, than recorded MIC for fungal test strains. Two fungal strains (Penicillium sp. and A. niger) show susceptible response to phenoloxidase using MgSO4 as inducer. Whereas, lysate-treated CaCl2 induced susceptibility only to A. niger. CONCLUSION: Hemocyanin shows better antimicrobial activity than phenoloxidase because of its broad-spectrum activity against bacterial and fungal strains tested. Hence, the hemocyanin may potentially become a new antimicrobial candidate to be discovered for a future use in treatment of resistant bacteria.

The relationship between the orientation of the glenoid and tears of the rotator cuff.

Our aim was to determine the most repeatable three-dimensional measurement of glenoid orientation and to compare it between shoulders with intact and torn rotator cuffs. Our null hypothesis was that glenoid orientation in the scapulae of shoulders with a full-thickness tear of the rotator cuff was the same as that in shoulders with an intact rotator cuff. We studied 24 shoulders in cadavers, 12 with an intact rotator cuff and 12 with a full-thickness tear. Two different observers used a three-dimensional digitising system to measure glenoid orientation in the scapular plane (ie glenoid inclination) using six different techniques. Glenoid version was also measured. The overall precision of the measurements revealed an error of less than 0.6 degrees. Intraobserver reliability (correlation coefficients of 0.990 and 0.984 for each observer) and interobserver reliability (correlation coefficient of 0.985) were highest for measurement of glenoid inclination based on the angle obtained from a line connecting the superior and inferior points of the glenoid and that connecting the most superior point of the glenoid and the most superior point on the body of the scapula. There were no differences in glenoid inclination (p = 0.34) or glenoid version (p = 0.12) in scapulae from shoulders with an intact rotator cuff and those with a full-thickness tear. Abnormal glenoid orientation was not present in shoulders with a torn rotator cuff.

Human fetal hemoglobin synthesis and its NH2-terminal acetylation in a rabbit reticulocyte lysate translational system.

The biosynthesis of the acetylated (Hb FIc) and the non-acetylated (Hb F0) human fetal hemoglobin components has been examined in a cell-free translational system. The poly(A)-RNA was isolated from umbilical cord blood samples and translated in the heterologous translational system derived from rabbit reticulocyte lysates in the presence of labeled amino acid(s) or acetyl-CoA. The amount of each hemoglobin or globin chain made in the system was determined by separating the synthesis products by cation-exchange chromatographic methods. The in vitro synthesis ratios were close to the FIc/Ftotal values of the respective hemolysates. The same conclusion could be reached by determining the specific activity ratios of Hb FIc/Hb F0. Co-migration of radioactivity peaks with absorbance peaks indicated the synthesis of that hemoglobin or globin chain. Confirmation of the synthesis of true gamma 0 and gamma Ic was accomplished by high-pressure liquid chromatographic separation of 3H-labeled tryptic peptides. Each peptide corresponded well with the radioactivity peak. Labeled acetyl-group incorporation into Hb FIc and gamma IcT-1 provided direct evidence for acetylation of gamma chains in Hb FIc. The data indicate that the mRNA itself dictates whether a protein is acetylated and, if so, to what extent. The control appears to be not unique to the human red cell system.

Influence of gamma-chain amino-terminal acetylation on subunit assembly of human fetal hemoglobin.

A human hemoglobin F subunit recombination study was performed to determine the relative efficiency of recombination of amino-terminally acetylated gamma-chains and non-acetylated chains with alpha-chains. The results of this work suggested that the acetylated gamma Ic-chains combined more readily with the alpha-chains than the non-acetylated gamma o-chains. An important factor in the function and assembly of multi-subunit macromolecules is the interaction of the unlike subunits. A model system for the study of such interactions has been the protein hemoglobin. With respect to the hemoglobin molecule, it has been noted that relative affinities of normal and mutant subunits for the unlike subunits can have a significant influence on hemoglobin synthesis at the post-translation level, i.e., subunit assembly [1,2]. A similar mechanism may control the formation of human Hb FIc, a hemoglobin with NH2-terminally acetylated gamma- (gamma Ic)-chains. In this instance acetylation may occur on the ribosomes before subunit assembly. A previous report from our laboratory showed a slight increase in the relative proportion of Hb FIc in cord blood samples with over 5% Hb Bart's [3]. The data were interpreted to be due to an influence of alpha-chain deficiency (alpha-thalassemia) on the formation of Hb FIc and Hb Fo tetramers by a preference of alpha-chains for gamma Ic-chains over gamma o- (non-acetylated gamma)-chains. The present study involves an examination of the relative affinity of the gamma Ic- and gamma o-chains for the normal alpha-chains in an in vitro recombination system.

Heterogeneity in the globin chain composition of a human minor fetal hemoglobin component.

The first hemoglobin found to contain an acetyl blocking group was the minor human fetal hemoglobin, Hb FI, present as 10-15% of the total fetal hemoglobin in umbilical cord blood red cells. Acetylation occurs at the amino-terminal glycine of the gamma-globin chain. Assays for the acetyl group by two different methods gave values less than the 2 per tetramer expected for a fully acetylated hemoglobin. We have purified acetylated fetal hemoglobin FIc to homogeneity. The globin chain composition of Hb FIc has been examined by both globin chain separation on CM-cellulose and by tryptic peptide mapping by HPLC. The identities of the gamma globin chains and of the gamma T-1 peptides were confirmed by amino acid analysis. Globin chain separation profiles showed the presence of 22.3 +/- 7.0% of gamma 0 globin (of the total gamma globin) in Hb FIc. Accordingly, the tryptic peptide maps of Hb FIc tetramers also showed the presence of a similar amount of gamma 0T-1 peptide. The gamma 0T-1 peptide was not present in the maps of isolated gamma Ic globin. It is evident that column purified Hb FIc contains a certain percentage of non-acetylated gamma-globin chains, thus indicating a hybrid globin chain composition for this minor fetal hemoglobin component.

Erythroid colony formation in vitro from the marrow of dogs with cyclic hematopoiesis: interrelationship of progenitor cells.

Committed erythroid progenitor cells (Colony Forming Units-Erythroid, CFU-E) have been studied in canine cyclic hematopoiesis (CH) utilizing a semi-solid methyl cellulose culture system. Erythroid colonies were stimulated by the addition of a standard volume of serum from normal dogs that had been phlebotomized and subjected to hypoxia. CFU-E fluctuated over the cycle in dogs with CH from concentrations 4--5 times normal during the periods of peripheral blood neutropenia to less than one tenth of normal during the phases of elevated peripheral blood neutrophil counts. In spite of these marked fluctuations there was no change in the proliferation rate of the CFU-E as estimated by the tritiated thymidine (3H-TdR) suicide technique. Failure to demonstrate a change in the CFU-E proliferation rate may be related to the relative maturity of these cells with the fluctuations in number resulting from a 'feed-in' from more immature cells. The results show that CFU-E fluctuate in the same phase as committed granulocytic progenitor cells (CFU-C). Our current knowledge of the interrelationships of marrow progenitor cells and events in the peripheral blood of dogs with CH is briefly reviewed and some additional questions, raised by recent studies regarding the pathogenesis of this disease, are discussed.

Tritiated thymidine incorporation into canine cyclic hematopoietic bone marrow in vitro.

To investigate changes in the proliferative activity of bone marrow cells in canine cyclic hematopoiesis, nonadherent cells were incubated for 1 h with tritiated thymidine either immediately after the cultures were established or following an 18-h preincubation period. The data suggest that changes in thymidine incorporation show a 12- to 14-day cycle that consists of two distinct phases. During the first six days of the cycle (from peripheral neutropenia to relative neutrophilia), two peaks of incorporation were observed. During the second phase (corresponding to the neutrophilia and oncoming neutropenia), thymidine incorporation was uniformly lower than control values. The change from an apparently cyclical process to a low stable value occurred after the wave of marrow myelopoiesis and close to a time point (days 8-10 of the cycle) at which we have recently suggested significant changes in cell release and/or proliferation take place. The data can be interpreted in the context of a periphery-to-stem-cell feedback loop through an intermediate cell population, probably of myeloid precursors.

Assessment of donor bone marrow cell-derived chimerism in transplantation tolerance using transgenic mice.

Allospecific skin graft prolongation can be induced in mice using antithymocyte globulin and allospecific donor bone marrow cells (DBMC). This enhancement may be due to the persistence of chimeric cells of donor origin in the host. In this study, we systematically investigated DBMC-derived chimerism in various lymphoid and nonlymphoid tissues over time. To do this, transgenic mice were used as a source of DBMC to clearly distinguish chimerism due only to the injected DBMC. Chimerism in various tissues was assessed at several times points after DBMC infusion by polymerase chain reaction amplification of tissue DNA using transgene specific primers. A cDNA probe specific for the transgene was used to demonstrate DBMC-derived chimerism in polymerase chain reaction products by the method of Southern. Although chimerism was initially detectable in most tissues tested 1 day after DBMC infusion, the presence of chimeric cells generally diminished over time. At 4 weeks or longer, chimerism was consistently confined to recipient skin. Furthermore, the chimeric cells in recipient skin persisted even after the allograft was rejected. In contrast to chimerism in recipient skin, chimerism became undetectable in donor skin as early as 2 weeks after DBMC infusion. The loss of chimerism in donor skin showed a temporal correlation with a reduction of chimerism in host bone marrow and lymphoid tissues that preceded rejection in all experiments by a range of 7-14 days. The use of DBMC from transgenic mice allowed a unique opportunity to monitor the kinetics of DBMC-derived chimerism. The presence of chimerism in the skin of mice in temporal association with chronic allograft rejection suggests that chimerism per se is not a reliable index of allogeneic unresponsiveness.

Promoter-region alleles of the TNF-alpha and IL-10 genes have no effect on pretransplant alloantibody production.

BACKGROUND: The presence of high levels of alloantibodies are known to be a risk factor in renal graft outcome. Expression level polymorphisms in cytokine genes are also thought to have an effect on allograft outcome, but the studies examining this have been inconsistent. This may be due to center-specific differences in immunosuppressive protocols. Therefore, we studied the effects of these polymorphisms on pretransplant class I alloantibody production in nonexogenously immunosuppressed candidates. METHODS: Tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) gene polymorphisms were assayed genotypically by PCR-SSP on 177 renal transplant candidates. Candidates with a peak goat antihuman immunoglobulin-enhanced T-cell panel reactive antibody (PRA) of >or=10% were considered to be positive for alloantibody (32% of 177 total). RESULTS: Previous transplants, transfusions, or pregnancies were all associated with alloantibody production, but TNF-alpha and IL-10 phenotypes were not. High levels of alloantibody production (peak PRA >50%) were also not effected by cytokine phenotype. CONCLUSIONS: These data suggest that differences in TNF-alpha and IL-10 phenotype do not effect a patient's likelihood of becoming sensitized by transfusions, pregnancies, and prior transplants.

Further studies of veto activity in rhesus monkey bone marrow in relation to allograft tolerance and chimerism.

Infusing the DR-/dim fraction of bone marrow cells (BMC) from an allogeneic kidney donor into rabbit antithymocyte globulin-treated transplant recipients delivers a tolerogenic signal, leading to functional allograft tolerance in rhesus monkeys without additional drug therapy. Our updated results in an expanded series show a median 131-day graft survival of recipients given DR-/dim donor BMC with a 23% 1-year survival (P < 0.00001 vs. rabbit antithymocyte globulin controls). Removing DRbright cells from donor BMC appeared to have a significant effect (P < 0.05). We have further investigated the tolerogenic mechanism within the experimental framework of the veto hypothesis in this preclinical model. In limiting dilution assays, we demonstrated the donor specificity of clonal inactivation of CTL precursors (CTLp) after in vitro or in vivo exposure to DR-/dim donor BMC, confirming specific tolerance. Additionally, in vitro studies confirmed the allogeneic specificity of CTLp inactivation in 3-cell MLR assays; minimal bystander effects were seen on normal CTLp responses to third party stimulator cells, while CTLp responses to the BMC donor's cells were abrogated in the same cultures. BMC mediating the veto effect were found to be resistant to L-leucyl-L-leucine methyl ester (Leu-leu-OMe), which excluded BMC-mediated cytotoxicity by NK or lymphokine-activated killer cells, CTL, or activated macrophages. In contrast, veto activity was abolished if the BMC were pretreated with either high dose UV-B light irradiation, mitomycin, or gamma-irradiation, indicating that BMC contained a UV-B-sensitive precursor of the veto effector, and that a proliferative step separated the two. Irradiation of DR-/dim donor BMC or administration of cyclophosphamide after infusion of nonirradiated BMC prevented the tolerogenic effect. Only recipients given nonirradiated DR-/dim donor BMC demonstrated PBL chimerism, which associated with functional deletion of antidonor CTLp and duration of graft survival. The Leu-leu-OMe resistance and the other properties of the allogeneic monkey CD3- CD2+ CD8+ BMC subpopulation that exhibits tolerance-promoting activity in vitro and in vivo lead us to postulate that a donor BMC-derived precursor population, possibly a dendritic cell population, may induce allogeneic unresponsiveness in this model.

TNF-alpha, IL-6, IFN-gamma, and IL-10 gene expression polymorphisms and the IL-4 receptor alpha-chain variant Q576R: effects on renal allograft outcome.

BACKGROUND: There has been much interest recently in the effects of various cytokine gene expression polymorphisms on graft outcome. However, the results of these investigations reveal the outcomes to be organ-specific and center-specific. We sought to confirm and add to some of the earlier findings by studying the impact of tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), interferon-gamma (IFN-gamma), and interleukin-6 (IL-6) polymorphisms and the interleukin-4 (IL-4) receptor alpha-chain variant on posttransplant renal allograft outcome. METHOD: TNF-alpha, IL-6, IFN-gamma, and IL-10 gene promoter region polymorphisms were assayed genotypically by PCR-SSP on 120 patients transplanted at the Albany Medical Center. These patients were also typed for the IL-4 receptor alpha-chain variant Q576R. RESULTS: Producers of high levels of the proinflammatory cytokine TNF-alpha were found to be at increased risk for acute rejection episodes if the allograft was mismatched for the molecular products of the class II region of the human major histocompatibility complex (HLA-DR). Expression level polymorphisms of the IL-6, IFN-gamma, and IL-10 genes were not associated with acute rejection episodes, nor was the IL-4 receptor alpha-chain variant Q576R. CONCLUSIONS: These data would suggest that the production of high levels of the cytokine TNF-alpha is especially detrimental to graft survival when the recipient's T-helper lymphocytes are being activated by mismatched donor HLA-class II antigens. Typing all potential kidney recipients for TNF-alpha, and providing well-matched organs for high producers of this cytokines, may be expected to increase rejection-free graft survival in these patients.

The facilitating effect of one-DR antigen sharing in renal allograft tolerance induced by donor bone marrow in rhesus monkeys.

Infusion of donor bone marrow cells (DBMC), a long-standing, successful strategy for inducing tolerance in experimental rodent transplantation models, can promote long-term acceptance of life-sustaining renal allografts in rhesus monkeys with no maintenance immunosuppression. To investigate the immunological basis for heterogeneity in duration of long-term graft acceptance following infusion of the DR-/dim fraction of DBMC into RATG-treated rhesus monkeys, we examined the relationship of recipient-donor major histo-compatibility class I and II DR matching to the development of antidonor antibody-dependent cellular cytotoxicity (ADCC) and renal allograft survival. The findings indicate a requirement for sharing one DR allele to achieve long-term graft acceptance. The observed immunological consequence of DR sharing that correlated with functional graft tolerance in this model was the suppression of early antidonor ADCC+ IgG antibody responses. Significant associations were observed between graft survival and suppression of ADCC antibody (P < 0.0005), graft survival and DR sharing (P < 0.005), and DR sharing and suppression of ADCC (P < 0.02). Early antidonor ADCC antibody responses associated with failure to maintain graft tolerance and were most consistently directed to donor class I. The required one DR antigen sharing in DBMC-induced suppression of antidonor class I antibody suggests a restriction for recipient DR, implying critical regulation of a response to donor antigen presented on recipient cells. We hypothesize a DBMC tolerogenic mechanism in which presentation of donor class I peptide by a shared DR allele configuration allows a veto effect by DBMC. Thus DR sharing would allow DBMC veto cells to reduce clonal expansion elicited by both the direct and indirect antigen presentation pathways.

Fast gel electrophoresis to analyze DNA-protein interactions.

A rapid method for electrophoresis of DNA-protein complexes is described. Popular "gel-shift" assays are performed using Pharmacia PhastSystem with its convenience of pre-cast gels and buffer strips and microprocessor-controlled high-resolution separation. Using this system, the products of a DNA binding reaction (DNA-protein complexes) can be separated from "free" DNA in less than one hour. DNA fragments as well as oligonucleotides have been used as targets with partially purified extracts containing sequence-specific DNA binding proteins. We present here a comparison of results of gel-shift assays obtained by conventional techniques and by our rapid "PhastShift" method which reduces the time, effort and technical expertise necessary to obtain reproducible results.

Ambient-temperature-stable molecular biology reagents.

We have processed biological materials to generate several reagents that are ambient temperature stable and ready to use. Stabilized biomolecules in a glassy matrix of carbohydrate polymers offer water-soluble reagents for complex molecular biology applications. This approach is particularly useful for reagent systems composed of enzymes, nucleotides and other components dispensed in single-use aliquots. Reconstitution of the glassy matrix delivers buffered enzymes and/or nucleotides for restriction, modification, sequencing and/or amplification of nucleic acids. These ambient-temperature-stable reagents allow a high level of reproducibility as they minimize the potential for pipetting errors. They also provide advantages in shipping, storage and subsequent handling. Added convenience includes elimination of setup time, cross contamination and refrigeration. Applications of ambient-temperature-stable biological reagents for routine molecular biology methods are presented.

Minimal criteria for the identification of Gardnerella vaginalis isolated from the vagina.

Vaginal swabs were examined for the presence of Gardnerella vaginalis. Of 294 isolates with appropriate colonial and cellular morphology subjected to an identification procedure, 203 (69%) were identified as G vaginalis. The 91 isolates not identified as G vaginalis were differentiated by their inability to ferment starch, cause diffuse beta haemolysis on human blood agar or hydrolyse hippurate. Other tests, often used in the identification of G vaginalis, were found to be insufficiently specific. Failure to ferment starch coexisted with failure to cause beta haemolysis and/or hydrolyse hippurate. The starch fermentation test may therefore be omitted. The tests for beta haemolysis and hippurate hydrolysis, being relatively simple to perform and interpret, are considered indispensable for the accurate identification of G vaginalis in the service laboratory.

Use of allogeneic bone marrow labeled with neomycin resistance gene to examine bone marrow-derived chimerism in experimental organ transplantation.

Posttransplant infusion of viable donor bone marrow cells (DBMC) has been shown in our previous studies to promote acceptance of incompatible kidney allografts in rhesus monkeys after treatment with polyclonal antithymocyte globulin to deplete peripheral T-lymphocytes. In this nonhuman primate model, the infusion of the DBMC is requisite for the induction of functional graft tolerance and specific MLR and CTLp unresponsiveness, although the relevant role and fate of bone marrow-derived chimeric cells is uncertain. Standard immunological and molecular techniques applied to this monkey model are unable to differentiate between chimeric cells derived from the infused DBMC and those derived from allograft-borne passenger leukocyte emigrants. To distinguish chimerism due to infused DBMC, we transduced DBMC with a functional neomycin resistance gene (Neo(r)) using the retroviral vector pHSG-Neo.Neo(r)-transduced BMC were infused into recipients approximately 2 wk after kidney transplantation and treatment with rabbit antithymocyte globulin. No maintenance immunosuppressive drugs were given. Genomic DNA isolated from peripheral blood leukocytes was used to monitor the presence ofNeo(r)-positive cells. Tissue samples obtained at necropsy also were assessed forNeo(r)-positive chimeric cells. The presence of DBMC-derived chimerism was assessed by polymerase chain reaction usingNeo(r) sequence-specific primers (PCR-SSP). Chimerism was detectable in recipient tissues at various times for up to 6 mo after DBMC infusion. These studies using gene transduction methodology indicate that a stable genetic marker can provide capability to examine DBMC-derived chimerism for prolonged periods in a nonhuman primate model. This approach should facilitate future studies in preclinical models to study the role and type of chimeric cell lineages in relation to functional allograft tolerance.

Gender and the comparison of self and observer ratings of anxiety and depression in adolescents.

OBJECTIVE: This study assessed the influence of gender on the comparability of self and observer ratings of anxiety and depression in adolescents. METHOD: Subjects were 75 inpatient adolescents who were administered structured interviews of the revised Hamilton Rating Scales for Depression (HRSD-R) and Anxiety (HARS-R) and read the Beck Depression Inventory (BDI) and Beck Anxiety Inventory (BAI). RESULTS: All measures demonstrated adequate internal consistency and validity. The correlation between the BDI and HRSD-R was significantly higher for females than males; of 11 symptoms that overlap on the BDI and HRSD-R, observers significantly agreed with males and females in their perceptions of 5 and 11 depressive symptoms, respectively. The correlation between the BAI and HARS-R did not differ significantly for males and females. CONCLUSIONS: Results suggest that self-reports of anxiety symptoms are a valid, cost-effective alternative to anxiety observer ratings for boys and girls' self-reports of depression are comparable to depression ratings by observers. There is the need to collect self-report information from adolescent boys because they may not communicate subjective symptoms of depression, e.g., guilt, to observers.