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1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) and the vitamin D receptor (VDR) have been reported to have an important role in the regulation of innate immunity. We earlier reported that the antimicrobial peptide cathelicidin is induced by 1,25(OH)2D3 in normal human bronchial epithelial cells with a resultant increase in antimicrobial activity against airway pathogens. In this study, we demonstrate that C/EBP alpha (C/EBPα) is a potent enhancer of human cathelicidin antimicrobial peptide (CAMP) gene transcription in human lung epithelial cells. In addition we found that C/EBPα functionally cooperates with VDR in the regulation of CAMP transcription. A C/EBP binding site was identified at -627/-619 within the CAMP promoter, adjacent to the vitamin D response element (VDRE; -615/-600). Mutation of this site markedly attenuated the transcriptional response to C/EBPα as well as to 1,25(OH)2D3, further indicating cooperation between these two factors in the regulation of CAMP. ChIP analysis using 1,25(OH)2D3 treated human lung epithelial cells showed C/EBPα and VDR binding to the CAMP promoter. C/EBPα has previously been reported to cooperate with Brahma (Brm), an ATPase that is component of the SWI/SNF chromatin remodeling complex. We found that dominant negative Brm significantly inhibited C/EBPα as well as 1,25(OH)2D3 mediated induction of CAMP transcription, suggesting the functional involvement of Brm. These findings define novel mechanisms involving C/EBPα, SWI/SNF, and 1,25(OH)2D3 in the regulation of CAMP in lung epithelial cells. These mechanisms of enhanced activation of the CAMP gene in lung epithelial cells suggest potential candidates for the development of modulators of innate immune responses for adjunct therapy in the treatment of airway infections., (© 2014 Wiley Periodicals, Inc.)

Attachment-Based Family Therapy (ABFT) is a manualized family-based intervention designed for working with depressed adolescents, including those at risk for suicide, and their families. It is an empirically informed and supported treatment. ABFT has its theoretical underpinnings in attachment theory and clinical roots in structural family therapy and emotion focused therapies. ABFT relies on a transactional model that aims to transform the quality of adolescent-parent attachment, as a means of providing the adolescent with a more secure relationship that can support them during challenging times generally, and the crises related to suicidal thinking and behavior, specifically. This article reviews: (1) the theoretical foundations of ABFT (attachment theory, models of emotional development); (2) the ABFT clinical model, including training and supervision factors; and (3) empirical support.

One in five adolescents in the United States has engaged in nonsuicidal self-injury (NSSI), one in eight have had serious thoughts of suicide, and one in 25 have attempted suicide. Research suggests that NSSI may increase risk for suicide attempt, yet little is known about the relationship between NSSI and suicidal ideation or attempts. In a primary care setting, 1,561 youth aged 14-24 years completed a brief, comprehensive, mental health screen as part of a routine well visit to determine which factors were most likely to predict suicidal ideation and attempt among youth engaging in NSSI. Results of recursive partitioning revealed that current depression and history of alcohol use best differentiated youth engaging in NSSI with low versus high risk for suicidal ideation and attempts. This simple algorithm is presented as a clinical screening tool that might aid medical providers in determining which youth would benefit from more intensive assessment and intervention., (© 2014 The American Association of Suicidology.)

There is a strong need for new broadly active antifungal agents for the treatment of oral candidiasis that not only are active against many species of Candida, including drug-resistant strains, but also evade microbial countermeasures which may lead to resistance. Host defense peptides (HDPs) can provide a foundation for the development of such agents. Toward this end, we have developed fully synthetic, small-molecule, nonpeptide mimetics of the HDPs that improve safety and other pharmaceutical properties. Here we describe the identification of several HDP mimetics that are broadly active against C. albicans and other species of Candida, rapidly fungicidal, and active against yeast and hyphal cultures and that exhibit low cytotoxicity for mammalian cells. Importantly, specificity for Candida over commensal bacteria was also evident, thereby minimizing potential damage to the endogenous microbiome which otherwise could favor fungal overgrowth. Three compounds were tested as topical agents in two different mouse models of oral candidiasis and were found to be highly active. Following single-dose administrations, total Candida burdens in tongues of infected animals were reduced up to three logs. These studies highlight the potential of HDP mimetics as a new tool in the antifungal arsenal for the treatment of oral candidiasis., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Antimicrobial peptides (AMPs) are found widely distributed through Nature, and participate in the innate host defense of each species. Fish are a great source of these peptides, as they express all of the major classes of AMPs, including defensins, cathelicidins, hepcidins, histone-derived peptides, and a fish-specific class of the cecropin family, called piscidins. As with other species, the fish peptides exhibit broad-spectrum antimicrobial activity, killing both fish and human pathogens. They are also immunomodulatory, and their genes are highly responsive to microbes and innate immuno-stimulatory molecules. Recent research has demonstrated that some of the unique properties of fish peptides, including their ability to act even in very high salt concentrations, make them good potential targets for development as therapeutic antimicrobials. Further, the stimulation of their gene expression by exogenous factors could be useful in preventing pathogenic microbes in aquaculture.

Friction melanosis is an acquired pigmentary disorder consisting of diffuse, brown pigmentation resulting from repeated rubbing of affected areas of the skin. An institutionalized child whose lower back was subjected to intense and repeated rocking displayed the characteristic lesions., (© 2012 Wiley Periodicals, Inc.)

This report summarizes the results of a study to develop an in vitro bioaccessibility (IVBA) extraction technique for estimating the relative bioavailability (RBA) of arsenic (As) in soil. The study was implemented in several steps. In step 1, key variables in the extraction protocol were identified. In step 2, 21 different extraction conditions were tested on 12 different soils with reliable RBA values measured in swine or monkeys to identify which yielded useful in vivo-in vitro correlations (IVIVC). In step 3, three extraction conditions were evaluated using 39 different test soils to make a final selection of the best IVIVC. In step 4, the within- and between-lab reproducibility of the extraction method was examined. The optimum IVIVC model for swine utilized a pH 1.5 IVBA extraction fluid, with an R (2) value of .723. For monkeys, the optimum IVIVC model was obtained using a pH 7 IVBA extraction fluid that contained phosphate, with an R (2) value of .755. Within-lab precision of IVBA results was typically less than 3%, with an average of 0.8% for all 4 labs. Between-lab variation in mean IVBA values was generally less than 7%, with an overall average of 3%. The principal advantages of this IVBA method compared to other in vitro methods described in the literature are that (1) the fluids and extraction conditions are simple, (2) the results have been calibrated against a larger data set than any other method, and (3) the method has been demonstrated to be reproducible both within and between labs.

Despite the well-documented association between history of sexual trauma (HSA) and suicide ideation, HSA is largely overlooked in suicide treatment studies. Existing studies showed that patients with a HSA have a weaker treatment response. In this randomized clinical trial for suicide ideation, HSA did not moderate treatment outcome for Attachment-Based Family Therapy (ABFT). Adolescents responded better to ABFT than a control condition, regardless of HSA status. At baseline, adolescents with HSA were also more likely to report past suicide attempts than those without HSA, indicating that they are a particularly important subgroup to consider when developing and evaluating interventions that target suicide ideation. Findings suggest that ABFT is a robust intervention for suicide ideation regardless of HSA., (PsycINFO Database Record (c) 2012 APA, all rights reserved.)

The airway epithelium plays a role in host defense through the binding of innate immune receptors, which leads to the activation of inflammatory mediators, including antimicrobial peptides. The active form of vitamin D, 1,25(OH)(2)D(3), induces the expression of the antimicrobial peptide LL-37 in both myeloid cells and airway epithelial cells (AEC). Here, we demonstrate that mRNA encoding triggering receptor expressed on myeloid cells (TREM)-1 was induced up to 12-fold by 1,25(OH)(2)D(3) in normal human bronchial epithelial (NHBE) cells and in well-differentiated cultures of six airway epithelial cell lines from patients with cystic fibrosis and healthy individuals. TREM-2 and DAP12 were also expressed in airway cultures, but not induced by vitamin D. Induction occurs through a vitamin D response element identified in its proximal promoter region, and was regulated by PU.1 expressed in the AEC. Activation of TREM-1 by a cross-linking antibody led to an induction of both human β-defensin-2 and TNF-α mRNA, demonstrating its functionality in these cells. Our results expand on the role played by the airway epithelium in innate immunity and suggest that vitamin D can modulate the innate immune defense of the airway epithelium, and could potentially be developed as an adjunctive therapy for airway infections.

Hepcidin is a cysteine-rich peptide involved in iron metabolism, inflammatory response and as antimicrobial peptide. Despite the fact that hepcidins have been identified in several fish species, only few have been completely characterized. This study, described the identification and complete molecular characterization of the hepcidin antimicrobial peptide 1 (HAMP1) gene of Alphestes immaculatus. Moreover, its specific expression level at both basal and lipopolysaccharide (LPS)-induced conditions in different tissues was also determined by real-time PCR. Results showed that the HAMP1gene consists of three exons and two introns encoding a preprohepcidin composed of 90 aa (24 aa for signal peptide, 40 aa for prodomain and 26 aa for mature peptide). The promoter region analysis revealed a TATA box sequence and several putative transcription factor binding sites. A comparative analysis showed CEBPα, CEBPβ, NF-kB, HNF3, GATA-1 and c-Rel as the most common found in fishes. The mature peptide possesses a pI of 8.34, which is the average among fish hepcidin. In addition, the structural modeling showed a hairpin structure with four putative disulfide bonds. A phylogenetic analysis revealed that this hepcidin gene is a HAMP1 class, and is clustered into the same group with the Serranid fish Epinephelus moara and the Antarctic fish Lycodichthys dearborni. Finally, the relative expression levels showed high basal values in liver and muscle, whereas in LPS-induced fish the relative expression tendency changed, with the highest values in spleen and head kidney tissues. This study describes the completely characterized HAMP1 gene of A. immaculatus and their patterns of expression level at different conditions and in different tissues, showing by first time muscle hepcidin expression could be relevant in the immune response in fish., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

hBD comprise a family of antimicrobial peptides that plays a role in bridging the innate and adaptive immune responses to infection. The expression of hBD-2 increases upon stimulation of numerous cell types with LPS and proinflammatory cytokines. In contrast, hBD-1 remains constitutively expressed in most cells in spite of cytokine or LPS stimulation; however, its presence in human PDC suggests it plays a role in viral host defense. To examine this, we characterized the expression of hBD-1 in innate immune cells in response to viral challenge. PDC and monocytes increased production of hBD-1 peptide and mRNA as early as 2 h following infection of purified cells and PBMCs with PR8, HSV-1, and Sendai virus. However, treatment of primary NHBE cells with influenza resulted in a 50% decrease in hBD-1 mRNA levels, as measured by qRT-PCR at 3 h following infection. A similar inhibition occurred with HSV-1 challenge of human gingival epithelial cells. Studies with HSV-1 showed that replication occurred in epithelial cells but not in PDC. Together, these results suggest that hBD-1 may play a role in preventing viral replication in immune cells. To test this, we infected C57BL/6 WT mice and mBD-1((-/-)) mice with mouse-adapted HK18 (300 PFU/mouse). mBD-1((-/-)) mice lost weight earlier and died sooner than WT mice (P=0.0276), suggesting that BD-1 plays a role in early innate immune responses against influenza in vivo. However, lung virus titers were equal between the two mouse strains. Histopathology showed a greater inflammatory influx in the lungs of mBD-1((-/-)) mice at Day 3 postinfection compared with WT C57BL/6 mice. The results suggest that BD-1 protects mice from influenza pathogenesis with a mechanism other than inhibition of viral replication.

Human gingival epithelial cells (GEC) produce peptides, such as β-defensins and the cathelicidin LL-37, that are both antimicrobial and that modulate the innate immune response. In myeloid and airway epithelial cells, the active form of vitamin D(3) [1,25(OH)(2)D(3)] increases the expression and antibacterial activity of LL-37. To examine the activity of vitamin D on the innate immune defense of the gingival epithelium, cultured epithelial cells were treated with either 10(-8) M 1,25(OH)(2)D(3) or ethanol for up to 24 h. A time-dependent induction of LL-37 mRNA up to 13-fold at 24 h in both standard monolayer and three-dimensional cultures was observed. Induction of the vitamin D receptor and the 1-α-hydroxylase genes was also observed. The hydroxylase was functional, as LL-37 induction was observed in response to stimulation by 25(OH)D(3). Through microarray analysis of other innate immune genes, CD14 expression increased 4-fold, and triggering receptor expressed on myeloid cells-1 (TREM-1) was upregulated 16-fold after 24 h of treatment with 1,25(OH)(2)D(3). TREM-1 is a pivotal amplifier of the innate immune response in macrophages, leading to increased production by inflammatory response genes. Activation of TREM-1 on the GEC led to an increase in interleukin-8 (IL-8) mRNA levels. Incubation of three-dimensional cultures with 1,25(OH)(2)D(3) led to an increase in antibacterial activity against the periodontal pathogen Aggregatibacter actinomycetemcomitans when the bacteria were added to the apical surface. This study is the first to demonstrate the effect of vitamin D on antibacterial defense of oral epithelial cells, suggesting that vitamin D(3) could be utilized to enhance the innate immune defense in the oral cavity.

This study examined whether improvement in parents' depression was linked with changes in their children's depressive symptoms and functioning. Participants were 223 parents and children ranging in age from 7 to 17 years old (M = 12.13, SD =2.31); 126 parents were in treatment for depression and 97 parents were nondepressed. Children were evaluated 6 times over 2 years. Changes in parents' depressive symptoms predicted changes in children's depressive symptoms over and above the effect of time; children's symptoms significantly predicted parents' symptoms. Trajectories of children's depressive symptoms differed significantly for children of remitted versus nonremitted depressed parents, and these differences were significantly predicted by their parents' level of depression. The relation between parents' and children's depressive symptoms was partially mediated by parental acceptance., (© 2011 The Authors. Child Development © 2011 Society for Research in Child Development, Inc.)

Unlabelled: Symptoms of internalizing disorders (depression, anxiety, somatic, trauma) are the major risk factors for suicide. Atypical suicide risk is characterized by people with few or no symptoms of internalizing disorders., Objective: In persons screened at intake to alcohol or other drug (AOD) treatment, this research examined whether person fit statistics would support an atypical subtype at high risk for suicide that did not present with typical depression and other internalizing disorders., Methods: Symptom profiles of the prototypical, typical, and atypical persons, as defined using fit statistics, were tested on 7408 persons entering AOD treatment using the Global Appraisal of Individual Needs (GAIN; Dennis et al., 2003a,b)., Results: Of those with suicide symptoms, the findings were as expected with the atypical group being higher on suicide and lower on symptoms of internalizing disorders. In addition, the atypical group was similar or lower on substance problems, symptoms of externalizing disorders, and crime and violence., Conclusions: Person fit statistics were useful in identifying persons with atypical suicide profiles and in enlightening aspects of existing theory concerning atypical suicidal ideation., (Copyright (c) 2009 Elsevier Ireland Ltd. All rights reserved.)

To more accurately assess the activity and role of epithelial cell-derived antimicrobial peptides in their native settings, it is essential to perform assays at the surfaces under relevant conditions. In order to carry this out, we utilize three-dimensional cultures of airway and gingival epithelium, which are grown at an air-liquid interface. Under these conditions, the cultures can be subjected to challenge with a variety of factors known to cause an increase in antimicrobial peptide gene expression. The functional relevance of this induction can then be assessed by quantifying antibacterial activity either directly on the surface of the cells or using the fluid secreted onto the apical surface of the cultures. The relative contribution of the peptides can also be measured by pre-incubation of the secreted fluid with specific inhibitory antibodies. Thus, a relatively inexpensive in vitro model can be used to evaluate the role of antimicrobial peptides in mucosal epithelium.

Background: Fetal alcohol spectrum disorder is a range of disabilities caused by gestational exposure to alcohol. FASD is the leading cause of preventable mental retardation and developmental disability in the United States, with an incidence of 1-10 per 1000 live births. FASD in Israel has yet to be examined systematically., Objectives: To evaluate professionals' experience, awareness and knowledge of FASD in Israel and their awareness of maternal consumption of alcohol, and to collect epidemiological data on the syndrome in Israel., Methods: A short questionnaire was sent to all 43 program directors of genetic institutes (n = 14) and child developmental centers in Israel (n = 29). Four questions related to their experience and knowledge of FASD. The epidemiological survey included data from all 17 hospitals in Israel and from the two main health management organizations within the public health care system., Results: The response rate was 98% (n = 42). A total of 38.1% of respondents reported having diagnosed at least one case of FASD and fewer than 10% of respondents stated that the knowledge regarding FASD among physicians in Israel was adequate. Developmental pediatricians were more likely to have diagnosed at least one case as compared to geneticists. During the period 1998-2007 the diagnosis of FASD appeared in the records of only 4 patients from the total number of 17 hospitals in Israel. During the same period only six patients were diagnosed at the HMO within the public health care system., Conclusions: Despite the accumulated knowledge on FASD in many countries and the increase in alcohol consumption in Israel, professionals' awareness of its potential damage is limited. Educational programs to increase physician awareness should accompany publicity campaigns warning the public of the dangers associated with alcohol consumption during pregnancy.

Objective: To evaluate the rate of depressive symptomatology and possible underlying factors in adoptive mothers during the transition to motherhood., Design: Cohort survey., Setting: General Community., Participants: Thirty-nine adoptive mothers of reproductive age registered with international adoption agencies., Interventions: All women completed the Edinburgh Postnatal Depression Scale (EPDS), the Beck Depression Inventory (BDI), and the Brief Symptom Inventory (BSI) before and 6 weeks after the adoption., Main Outcome Measures: Responses were compared between the study group and published findings for biological mothers in the general population, and within the study group, before and after adoption., Results: Symptoms of depression were found in 15.4% of the study group. This rate was similar to that for postpartum depression in the general population, and lower than the rate recorded in the study group before adoption (25.6%). All women with symptoms of depression after the adoption had also shown evidence of depressive features before the adoption. Similar findings were noted for other psychopathologies as well., Conclusion: Adopting a child does not cause new-onset, reactive depression among adoptive mothers. It may even lead to a decrease in depressive features, perhaps in response to relief from other adjustment difficulties.

Background: Studies from many countries have reported an increasing prevalence of autistic spectrum disorder in childhood. No comprehensive epidemiological studies of ASD have been performed in Israel., Objectives: To describe time trends in the reported number of patients with ASD in Israel and to characterize the demographic features of the reported patients., Methods: We reviewed the charts of the National Insurance Institute of Israel from 1972 to 2004 for all children with a diagnosis of ASD receiving disability benefits., Results: A total of 3509 children met the study criteria. Eighty percent were boys and 98% were Jewish. The incidence data showed an increase in the number of cases from zero in 1982-84 and 2 (1.2 per million capita under 18 years) in 1985 to a high of 428 cases in 2004 (190 per million)., Conclusions: This is the first comprehensive study of the incidence of ASD in Israel. According to data derived from official health records, the rate of occurrence of ASD has substantially increased in the last 20 years. Further studies are needed to determine if this is a true increase or if the findings were confounded by external factors, such as recent improvements in diagnostic measures and social stigmas.

Antimicrobial peptides (AMPs) are multi-functional peptides whose fundamental biological role in vivo has been proposed to be the elimination of pathogenic microorganisms, including Gram-positive and -negative bacteria, fungi, and viruses. Genes encoding these peptides are expressed in a variety of cells in the host, including circulating phagocytic cells and mucosal epithelial cells, demonstrating a wide range of utility in the innate immune system. Expression of these genes is tightly regulated; they are induced by pathogens and cytokines as part of the host defense response, and they can be suppressed by bacterial virulence factors and environmental factors which can lead to increased susceptibility to infection. New research has also cast light on alternative functionalities, including immunomodulatory activities, which are related to their unique structural characteristics. These peptides represent not only an important component of innate host defense against microbial colonization and a link between innate and adaptive immunity, but also form a foundation for the development of new therapeutic agents.

Parental negativity is associated with the onset and maintenance of adolescent depression. Reducing parental negativity is a primary focus of family-based treatments for this clinical population. This study examined the association between therapist relationship-facilitating and attachment-oriented interventions and the valence (i.e., positivity-negativity) of parents' attitudes toward their depressed adolescent in a sample of 13 sessions of attachment-based family therapy. Lag sequential analyses revealed that in good alliance sessions relationship-facilitating interventions, such as empathy and positive regard for the parent, were associated with parents' nonnegative attitudes toward their adolescent in the five speech turns subsequent to the intervention. Attachment-oriented interventions, such as relational reframes, addressing core relational themes, and highlighting vulnerable emotions, were also intermittently associated with nonnegative parental attitudes in good alliance sessions. No such effects were evident for the comparison interventions. This study represents a first step in the process of testing specific strategies for reducing parental negativity in family therapy.

Environmental factors, such as nutritional status, physical activity, and drug therapy, can affect bone mineralization. Our objective was to evaluate the relationship between nutritional status, physical activity, and bone mineralization as assessed by multisite quantitative ultrasound technology in children. The study group comprised 67 children, aged 6-17 years (mean, 9.4), attending a primary care clinic. Data on calcium intake and physical activity were collected using a detailed questionnaire. Speed of sound measurements were performed at the distal 1/3 radius and the midshaft tibia using Sunlight Omnisense apparatus. The reported mean calcium intake was 1105 mg/day. There was a significant difference in Z-scores at the radius and tibia between the low-and high-calcium-intake groups (P = 0.004, P = 0.035, respectively). A similar difference was found between the low-and normal-physical-activity groups (P = 0.015, P = 0.036, respectively). In this pilot study, a positive association was found between calcium intake, physical activity, and bone status, as assessed by the quantitative ultrasound technique.

Antimicrobial peptides (AMPs) are generally produced as precursor peptides containing a signal sequence, a pro-region and the mature peptide. A computational analysis of beta-defensin precursors predicts cleavage solely by signal peptidase to release the mature peptide, with no pro-region. This supports the extensive transcriptional control of beta-defensin expression compared with other AMP genes.

Background: Antimicrobial peptides (AMPs) such as cathelicidins contribute to initial defense of the airway against inhaled pathogens. Recent studies have shown that the hormonally active form of vitamin D(3), 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) up-regulates AMP gene expression in several established cell lines. Furthermore, serum levels of vitamin D are often deficient in cystic fibrosis (CF) patients., Methods: We investigated the effect of 1,25(OH)(2)D(3) on AMP mRNA levels in primary cultures of normal human bronchial epithelial (NHBE) cells by real-time PCR, and protein levels by Western blot. Antimicrobial activity of airway surface fluid from these cells was measured by in vitro assay against laboratory strains of bacteria., Results: Treatment of NHBE cells with 1,25(OH)(2)D(3) (10(-8)M), resulted in a 10-fold up-regulation of cathelicidin mRNA levels after 12 h, which was augmented 2-fold with co-incubation of 1 mM Calcium. Moreover, 1,25(OH)(2)D(3) induced antimicrobial activity against the airway pathogens Bordetella bronchiseptica and Pseudomonas aeruginosa. 1,25(OH)(2)D(3) induced cathelicidin mRNA expression equally in both normal and CF bronchial epithelial cells., Conclusions: Elucidation of the effect of 1,25(OH)(2)D(3) on cathelicidin expression in NHBE cells and CF bronchial epithelial cells will aid in the development of novel therapeutic agents for treatment of airway infections in CF.

Antimicrobial peptides (AMPs) are naturally occurring, broad-spectrum antimicrobial agents that have recently been examined for their utility as therapeutic antibiotics. Unfortunately, they are expensive to produce and are often sensitive to protease digestion. To address this problem, we have examined the activity of a peptide mimetic whose design was based on the structure of magainin, exhibiting its amphiphilic structure. We demonstrate that this compound, meta-phenylene ethynylene (mPE), exhibits antimicrobial activity at nanomolar concentrations against a variety of bacterial and Candida species found in oral infections. Since Streptococcus mutans, an etiological agent of dental caries, colonizes the tooth surface and forms a biofilm, we quantified the activity of this compound against S. mutans growing under conditions that favor biofilm formation. Our results indicate that mPE can prevent the formation of a biofilm at nanomolar concentrations. Incubation with 5 nM mPE prevents further growth of the biofilm, and 100 nM mPE reduces viable bacteria in the biofilm by 3 logs. Structure-function analyses suggest that mPE inhibits the bioactivity of lipopolysaccharide and binds DNA at equimolar ratios, suggesting that it may act both as a membrane-active molecule, similar to magainin, and as an intracellular antibiotic, similar to other AMPs. We conclude that mPE and similar molecules display great potential for development as therapeutic antimicrobials.

Background: Developmental and behavioral pediatrics has emerged as an area of special interest and new responsibility for pediatricians. The aim of this study was to evaluate the impact of training, experience, and other factors on pediatricians' satisfaction with their abilities to care for children with developmental, behavioral and psychosocial problems., Methods: A questionnaire was sent to 211 pediatricians working in primary care clinics in the community in Israel. Items included personal characteristics and experience, practice and satisfaction with the care of children with developmental, behavioral and psychosocial problems. Overall, pediatrician satisfaction with their personal abilities in this domain was defined as the dependent variable on multivariate analysis., Results: The response rate was 76.3% (n = 161). Pediatricians' satisfaction with their overall professional ability to care for children with developmental, behavioral and psychosocial problems was significantly and positively associated with the following independent variables: (i) satisfaction with training received in the child development field; (ii) satisfaction with psychiatric updates; (iii) general satisfaction with the available child development services; (iv) prescription of methylphenidate to children with attention deficit/hyperactivity disorder; and (v) having completed their medical studies in a country in which the issue is emphasized., Conclusions: Imparting more knowledge and skills in child development and behavioral pediatrics in pediatric residency and continuing education programs will help improve the attitudes of primary care pediatricians towards developmental and behavioral problems, encourage them to treat these patients and their families, and better the quality of care.

Sequencing of the 300 bp region of the mitochondrial cytochrome b gene was done. Genetic analysis was carried out for the first time in three exotic breeds (Giant White, Soviet Chinchilla, and German Angora) of European rabbit (Oryctolagus cuniculus) to determine intra- and interspecific variability and to measure the genetic distance. The frequencies of types of mutations (transition, transversion, deletion, and insertion) were also determined. This study throws light on matrilineage of breeds that arise due to interbreed crosses and the genetic management of a stocked rabbit breeding population.

Background and Objective: Human beta-defensins have been identified in the oral cavity and are predicted to play a role in the defense against pathogenic bacteria. Homologous rat beta-defensins (RBDs) have been identified, but their expression in the oral cavity has not been examined. Therefore, the aim of this study was to investigate the expression of innate immune mediators in the rat gingival epithelium., Material and Methods: Rats were pretreated with antibiotics to depress the normal oral flora, followed by the introduction of Actinobacillus actinomycetemcomitans in their food to allow colonization and the development of periodontal disease. At various time points, animals were killed and the gingival epithelium was extracted. Semiquantitative reverse transcription-polymerase chain reaction was performed to measure RBD and Toll-like receptor (TLR) mRNA levels., Results: Three beta-defensins (RBD-1, -2 and -5) and two TLRs (TLR-3 and -4) are expressed in normal rat gingival epithelium. After the introduction of A. actinomycetemcomitans, RBD-1 and RBD-2 mRNA levels increased for the first week followed by a return to basal levels. No change in TLR mRNA levels was observed., Conclusion: The rat model provides a good system for experimental analysis of the innate immune response to periopathogenic bacteria in the oral cavity, as well as the potential role of beta-defensins in the host response to colonization.

Poor ambient air quality is associated with increased morbidity and mortality, including respiratory infections. However, its effects on various host-defense mechanisms are poorly understood. This study utilized an in vitro model to study the effect of particulate matter (PM(2.5)) on one antimicrobial mechanism of host defense in the airway, beta-defensin-2 and its bovine homologue, tracheal antimicrobial peptide (TAP) induction in response to lipopolysaccharide (LPS) and IL-1beta. Our model utilized cultured primary bovine tracheal epithelial (BTE) cells and the human alveolar type II epithelial cell line, A549, treated with 0-20 microg/cm(2) residual oil fly ash (ROFA) for 6 h. The cells were then washed and stimulated for 18 h with 100 ng/ml LPS or for 6 h with 100 ng/ml IL-1beta. ROFA inhibited the LPS-induced increase in TAP mRNA and protein without inducing significant cytotoxicity. As little as 2.5 microg/cm(2) of ROFA inhibited LPS-induced TAP gene expression by 30%. The inhibitory activity was associated with the soluble fraction and not the washed particle. The activity in the leachate was attributed to vanadium, but not nickel or iron. SiO(2) and TiO(2) were utilized as controls and did not inhibit LPS induction of TAP gene expression in BTE. ROFA also inhibited the increase of IL-1beta-induced human beta-defensin-2, a homologue of TAP, in A549 cells. The results show that ROFA, V(2)O(5), and VOSO(4) inhibit the ability of airway epithelial cells to respond to inflammatory stimuli at low, physiologically relevant doses and suggest that exposure to these agents could result in an impairment of defense against airborne pathogens.

The airway provides numerous defense mechanisms to prevent microbial colonization by the large numbers of bacteria and viruses present in ambient air. An important component of this defense is the antimicrobial peptides and proteins present in the airway surface fluid (ASF), the mucin-rich fluid covering the respiratory epithelium. These include larger proteins such as lysozyme and lactoferrin, as well as the cationic defensin and cathelicidin peptides. While some of these peptides, such as human beta-defensin (hBD)-1, are present constitutively, others, including hBD2 and -3 are inducible in response to bacterial recognition by Toll-like receptor-mediated pathways. These peptides can act as microbicides in the ASF, but also exhibit other activities, including potent chemotactic activity for cells of the innate and adaptive immune systems, suggesting they play a complex role in the host defense of the airway. Inhibition of antimicrobial peptide activity or gene expression can result in increased susceptibility to infections. This has been observed with cystic fibrosis (CF), where the CF phenotype leads to reduced antimicrobial capacity of peptides in the airway. Pathogenic virulence factors can inhibit defensin gene expression, as can environmental factors such as air pollution. Such an interference can result in infections by airway-specific pathogens including Bordetella bronchiseptica, Mycobacterium tuberculosis, and influenza virus. Research into the modulation of peptide gene expression in animal models, as well as the optimization of peptide-based therapeutics shows promise for the treatment and prevention of airway infectious diseases.

Objective: To provide an update on the state of the art of family-based treatment research., Method: Randomized clinical trials conducted in the past 10 years that included parents as a primary participant in treatment of child and adolescent psychiatric problems were reviewed. Studies were identified from major literature search engines (e.g., PsycINFO, Medline). Current significant pilot work was identified in the National Institute of Mental Health Computer Retrieval of Information on Scientific Projects (CRISP) Web page or from the authors themselves., Results: Family treatments have proven effective with externalizing disorders, particularly conduct and substance abuse disorders, and in reducing the comorbid family and school behavior problems associated with attention-deficit/hyperactivity disorder. Several new studies suggest that family treatments or treatment augmented by family treatments are effective for depression and anxiety., Conclusions: For many disorders, family treatments can be an effective stand-alone intervention or an augmentation to other treatments. Engaging parents in the treatment process and reducing the toxicity of a negative family environment can contribute to better treatment engagement, retention, compliance, effectiveness, and maintenance of gains. Recommendations for the next decade of research and some implications of family-based treatment for child and adolescent psychiatry are explored.

To determine the role of human beta-defensin 2 (HBD-2) in human tuberculosis, we studied the in vitro induction of HBD-2 gene expression by Mycobacterium tuberculosis H37Rv infection in the human lung epithelial cell line A549, in alveolar macrophages (AM), and in blood monocytes (MN) by reverse transcription-PCR. We also studied the induction of HBD-2 gene expression by mannose lipoarabinomannan (manLAM) from M. tuberculosis. Intracellular production of HBD-2 peptide was detected by immunocytochemistry and electron microscopy. Our results demonstrated that there was induction of HBD-2 mRNA in A549 cells after infection with M. tuberculosis at various multiplicities of infection (MOI) and that there was stimulation with manLAM. AM expressed the HBD-2 gene only at a high MOI with M. tuberculosis. MN did not express HBD-2 at any of the experimental M. tuberculosis MOI. Immunostaining revealed the presence of intracellular HBD-2 peptide in A549 cells following infection with M. tuberculosis, and the staining was more intense in areas where there were M. tuberculosis clusters. By using electron microscopy we also demonstrated production of HBD-2 after M. tuberculosis infection and adherence of HBD-2 to the membranes of M. tuberculosis. Alveolar epithelial cells are among the first cells to encounter M. tuberculosis following aerogenic infection. As HBD-2 has been shown to control growth of M. tuberculosis and has chemotactic activity, our results suggest that HBD-2 induction by M. tuberculosis may have a role in the pathogenesis of human tuberculosis.

Defensins are key participants in mucosal innate defense. The varied antimicrobial activity and differential distribution of defensins at mucosal sites indicate that peptide repertoires are tailored to site-specific innate defense requirements. Nonetheless, few studies have investigated changes in peptide profiles and function after in vivo pathogen challenge. Here, we determined defensin profiles in urethral secretions of healthy men and men with Chlamydia trachomatis- and Neisseria gonorrhoeae-mediated urethritis by immunoblotting for the epithelial defensins HBD1, HBD2, and HD5 and the neutrophil defensins HNP1 to -3 (HNP1-3). HBD1 was not detectable in secretions, and HBD2 was only induced in a small proportion of the urethritis patients; however, HD5 and HNP1-3 were increased in C. trachomatis infection and significantly elevated in N. gonorrhoeae infection. When HNP1-3 levels were low, HD5 appeared mostly as the propeptide; however, when HNP1-3 levels were >10 microg/ml, HD5 was proteolytically processed, suggesting neutrophil proteases might contribute to HD5 processing. HD5 and HNP1-3 were bactericidal against C. trachomatis and N. gonorrhoeae, but HD5 activity was dependent upon N-terminal processing of the peptide. In vitro proteolysis of proHD5 by neutrophil proteases and analysis of urethral secretions by surface-enhanced laser desorption ionization substantiated that neutrophils contribute the key convertases for proHD5 in the urethra during these infections. This contrasts with the small intestine, where Paneth cells secrete both proHD5 and its processing enzyme, trypsin. In conclusion, we describe a unique defensin expression repertoire in response to inflammatory sexually transmitted infections and a novel host defense mechanism wherein epithelial cells collaborate with neutrophils to establish an antimicrobial barrier during infection.

Expression of innate immune genes such as beta-defensins is induced in airway epithelium by bacterial components via activation of NF-kappaB. We show here that live Gram-negative bacteria can similarly stimulate this pathway, resulting in upregulation of the beta-defensin tracheal antimicrobial peptide (TAP) in primary cultures of bovine tracheal epithelial cells (TECs), by a Toll-like receptor 4 (TLR4)-mediated pathway. The Gram-negative airway pathogen Bordetella bronchiseptica possesses a type III secretion system previously suggested to inhibit the nuclear translocation of NF-kappaB in a cell line by immunohistochemistry. We therefore hypothesized that this pathogen might interfere in the innate immune response of the epithelium. Exposure of TECs to wild-type B. bronchiseptica suppressed the activation of NF-kappaB and the subsequent induction of TAP mRNA levels, whereas a type III secretion-defective strain did not. These results suggest a mechanism for bacterial evasion of the innate immune response in the airway, which could allow for the observed persistent colonization of this pathogen.

This research utilized the Pichia pastoris expression system for recombinant expression of cDNA of pleurocidin, a small (2.7 kd) antimicrobial peptide isolated from winter flounder (Pleuronectes americanus). The Pichia vector contains the alcohol oxidase gene promoter (AOX 1), which under the induction of methanol allows for expression of heterologous protein gene inserted downstream in the vector. Two strains of P pastoris were used as host cells, the wild type (P pastoris X-33((mut(+)))) and the mutant (P pasatoris KM71H((mut(s))) ). Polymerase chain reaction (PCR) and DNA sequencing showed that pleurocidin cDNA was successfully integrated into the P pastoris genome. Reverse transcription (RT)-PCR showed that pleurocidin was transcribed by both Pichia host strains. Affinity chromatography, SDS-PAGE, and immunological techniques were used for purification and detection of recombinant peptide. Although there was strong evidence of transcription of pleurocidin cDNA, the Pichia system requires further optimization to obtain detectable levels of this small peptide.

This article presents the main outcome findings from two inter-related randomized trials conducted at four sites to evaluate the effectiveness and cost-effectiveness of five short-term outpatient interventions for adolescents with cannabis use disorders. Trial 1 compared five sessions of Motivational Enhancement Therapy plus Cognitive Behavioral Therapy (MET/CBT) with a 12-session regimen of MET and CBT (MET/CBT12) and another that included family education and therapy components (Family Support Network [FSN]). Trial II compared the five-session MET/CBT with the Adolescent Community Reinforcement Approach (ACRA) and Multidimensional Family Therapy (MDFT). The 600 cannabis users were predominately white males, aged 15-16. All five CYT interventions demonstrated significant pre-post treatment during the 12 months after random assignment to a treatment intervention in the two main outcomes: days of abstinence and the percent of adolescents in recovery (no use or abuse/dependence problems and living in the community). Overall, the clinical outcomes were very similar across sites and conditions; however, after controlling for initial severity, the most cost-effective interventions were MET/CBT5 and MET/CBT12 in Trial 1 and ACRA and MET/CBT5 in Trial 2. It is possible that the similar results occurred because outcomes were driven more by general factors beyond the treatment approaches tested in this study; or because of shared, general helping factors across therapies that help these teens attend to and decrease their connection to cannabis and alcohol.

A PBPK model for 2,4-D was developed that involves flow-limited pH trapping modified to consider tissue binding, binding to plasma, and high-dose inhibition of urinary excretion. The PBPK model provides reasonable estimates of the kinetics of 2,4-D in rats as well as in humans, providing a common metric for expressing risk. The risk characterization for 2,4-D based on the PBPK model is consistent with that based on standard risk assessment methods, except that the apparent variability in the risk characterization is reduced. The model demonstrates that non-linear pharmacokinetics and inhibition of urinary excretion would not be expected in occupational exposures. This case study suggests that preliminary PBPK models could be developed for numerous pesticides based on commonly available data. If properly validated with well-designed worker exposure studies, such models may be useful in more complete assessments of risks to workers as well as members of the general public.

Background: The prevalence rates of pervasive developmental disorder (PDD) have risen in the West over the last 10 years. There is argument over the etiology of this change in rates. Social and cultural processes including migration have been hypothesized. Israel, as a country of ongoing immigration with a national registry of children diagnosed with PDD, offers an opportunity to compare rates of PDD among immigrants from developing countries and native Israelis., Method: A Social Security national registry of 1004 children diagnosed with PDD was reviewed and rates were calculated using data extracted from the Israel National Bureau of Statistics. Of all Jewish children that were born in the years 1983-1997 and who are currently living in Israel, we defined four groups: (1). native Israelis of non-Ethiopian extraction (N = 1198, 300), (2). native Israelis of Ethiopian extraction (N = 15600), (3). immigrants of non-Ethiopian extraction (N = 110300) and (4). children born in Ethiopia (N = 11800). A further breakdown of groups 1 and 3 by well-characterized ethnic or geographical origins was not possible., Results: The rate of PDD was significantly elevated in native Israelis as compared to all immigrant children. Among immigrants, the rate of PDD in Ethiopian-born children was lower than that of those born in other countries. The rate of PDD in immigrant Ethiopian children was much lower than in native Israeli children of Ethiopian extraction., Conclusions: Birth in Israel, an industrialized country, is a marker for an environmental risk factor for PDD. This may indicate that gestation, birth or infancy in industrialized countries exposes children to environmental insults that increase the risk for contracting PDD.

Production of human beta-defensin1 (HBD1) in response to LPS in monocytes, myeloid dendritic cells and plasmacytoid dendritic cells (PDC) was examined. Since PDC make up only 0.1-0.5% of the peripheral blood mononuclear cell population, we developed a method to determine HBD1 peptide levels using four-color flow cytometry, which can examine several cell surface or intracellular markers at once. Coupled with intracellular flow cytometry, we determined that PDC and monocytes only made significant amounts of HBD1 when exposed to >50ng/ml LPS for 2h. This response was limited to monocytes when ultrapure LPS was used, and was inhibited in PDC by chloroquine treatment.

The iron-binding protein lactoferrin is a ubiquitous and abundant constituent of human exocrine secretions. Lactoferrin inhibits bacterial growth by sequestering essential iron and also exhibits non-iron-dependent antibacterial, antifungal, antiviral, antitumor, anti-inflammatory, and immunoregulatory activities. All of these non-iron-dependent activities are mediated by the highly charged N terminus of lactoferrin. In this study we characterized a Lys/Arg polymorphism at position 29 in the N-terminal region of human lactoferrin that results from a single nucleotide polymorphism in exon 1 of the human lactoferrin gene. We expressed cDNAs encoding both lactoferrin variants in insect cells and purified the two proteins by ion exchange chromatography. The two lactoferrin variants exhibited nearly identical iron-binding and iron-releasing activities and equivalent bactericidal activities against a strain of the gram-negative bacterium Actinobacillus actinomycetemcomitans. When tested against the gram-positive species Streptococcus mutans and Streptococcus mitis, however, lactoferrin containing Lys at position 29 exhibited significantly greater bactericidal activity than did lactoferrin containing Arg. In addition, the Lys-containing lactoferrin stimulated bovine tracheal epithelial cells to synthesize much higher levels of tracheal antimicrobial peptide mRNA than did the Arg-containing variant. A genotyping assay that distinguished between the two alleles based on a polymorphic EarI restriction site showed that the Lys and Arg alleles had frequencies of 24% and 76%, respectively, among 17 healthy human subjects, and 72% and 28%, respectively, among nine patients with localized juvenile periodontitis. Our findings suggest that these two lactoferrin variants are functionally different and that these differences may contribute to the pathogenesis of localized juvenile periodontitis.

More than 85% of peak skeletal mass is accrued by the age of 18 years, making bone growth during childhood and adolescence a critical process. The purpose of this study is to establish pediatric reference curves for bone Speed of Sound (SOS) as measured by multi-site quantitative ultrasound. Analysis was performed on a total of 1085 healthy subjects ages 0-18 years (595 females, 490 males). Demographic and anthropometric parameters (height and weight), as well as data on calcium intake and physical activity, were collected. Ultrasound bone measurements were performed at the mid-shaft tibia and the distal third of the radius (Sunlight Omnisense 7000P). An age-related speed of sound (SOS) curve that describes SOS changes at the tibia and radius in both genders was demonstrated. SOS showed a steep increase during the first 5 years of life in both genders at the tibia and radius. The period between the ages of 6-11 years is characterized with a very shallow increase in SOS at both sites. Thereafter, during the pubertal period, there is a second growth burst in SOS, starting at age 11 for girls and age 14 for boys. No significant meaningful correlation was found between the anthropometric parameters (height, weight and BMI) and the SOS measurements after the age parameter was controlled. Subjects who reported low physical activity levels were found to have lower Z-scores than their counterparts (P<0.05). The SOS of pre-menarche girls was significantly lower than that of post-menarche girls at the radius and tibia (P<0.05). The level of calcium intake did not correlate with bone SOS. Intra-operator precision measurements were 0.36% (0.25-0.47%) at radius and 0.30% (0.20-0.40%) at the tibia. To date, there is no widely accepted classification or clinical working guidelines for children's bone health assessment or prediction of fracture risk based on bone strength measurements in children. This study establishes a pediatric reference curve for the Omnisense, and therefore supports the feasibility of using Sunlight Omnisense 7000P, a multi-site bone sonometer, for the assessment of pediatric bone properties. Further studies mainly in different diseased children groups should further support the use of such a basic tool for clinical evaluation, assisting the physician to work towards healthy bones for his patients.

With the increase in the number of children being adopted by Israeli families from abroad, the medical system has to face heretofore unfamiliar medical and developmental challenges. Many of the biological mothers of these children were substance abusers and suffered from a variety of illnesses and nutritional deficiencies during their pregnancy. Their infants and young children were placed in orphanages at a very young age. The ensuing emotional and environmental deprivation in many cases impacted negatively on their neurological, physical and emotional development. Whereas our previous survey dealt with the purely medical aspects of international adoption, the current review deals with the growth and developmental issues involved in raising these children.

Few effective psychosocial treatment models for depressed adolescents have been developed, let alone ones that use the developmentally potent context of the family as the focus of intervention. Attachment-based family therapy (ABFT) is a brief, manualized treatment model tailored to the specific needs of depressed adolescents and their families. Attachment theory serves as the main theoretical framework to guide the process of repairing relational ruptures and rebuilding trustworthy relationships. Empirically supported risk factors for depression are the primary problem states that therapists target with specific treatment strategies or tasks. Parent problem states include criticism/hostility, personal distress, parenting skills, and disengagement. Adolescent problem states include motivation, negative self-concept, poor affect regulation, and disengagement. Intervention tasks include relational reframing, building alliances with the adolescent and with the parent, addressing attachment failures, and building competency. A small, randomized clinical trial provides initial support for the model. Several process research studies, using both qualitative and quantitative methods, have helped refine the clinical guidelines for each treatment task. ABFT is a promising new treatment for depressed adolescents and more research on it is warranted.

Lung tissue removed from neonatal calves with acute Mannheimia haemolytica pneumonia showed that rapid up-regulation of the basal mRNA expression of tracheal antimicrobial peptide (TAP), NF-kappa B, and intercellular adhesion molecule 1 occurred after infection; TAP and interleukin 8 expression were highly correlated. This work suggests that the coordinated expression of beta-defensin and inflammatory elements occurs during bacterial pneumonia.

The five manual-guided treatment models tested in the Cannabis Youth Treatment study funded by the Center for Substance Abuse Treatment are described. The five models include (a) a 6-week intervention consisting of two sessions of individual motivational enhancement therapy plus three sessions of group cognitive behavioral therapy (MET/CBT5); (b) a 12-week intervention consisting of two sessions of motivational enhancement therapy plus 10 sessions of group cognitive behavioral therapy treatment (MET/CBT12); (c) a 12-week intervention consisting of MET/CBT12 plus the family support network (FSN), a multi-component intervention that includes parent education, family therapy and case management; (d) a 12-week intervention based on the adolescent community reinforcement approach (ACRA), an individual behavioral treatment approach designed to help adolescents and their parents reshape their environment and learn new skills; and (e) multi-dimensional family therapy (MDFT), a multi-faceted, developmentally and contextually oriented family-based model targeting individual, family and social systems. For each model, we describe the treatment background and/or its empirical support, its theoretical underpinnings, its goals and proposed treatment mechanism and the structure and content of each treatment. Procedures used for maintaining treatment fidelity and monitoring quality assurance are also described. These interventions represent the first readily available, manual-guided interventions to be evaluated in a large randomized field study for this population. Consequently, these manuals have the potential to advance treatment and research for adolescents with substance use disorders.

Aims: Despite recent advances in the economic evaluation of adult substance abuse treatment, information and basic research is lacking on the cost of adolescent substance abuse treatment. The present study conducted an economic cost analysis of several outpatient adolescent treatment approaches., Design: The Cannabis Youth Treatment (CYT) study evaluated five structured treatments for cannabis-using adolescents. One of the approaches was implemented by all of the four geographically and institutionally diverse treatment facilities collaborating in CYT; each of the other four approaches was implemented in two of the sites. Using the Drug Abuse Treatment Cost Analysis Program (DATCAP), the economic cost of each site-specific treatment was determined., Findings: The average economic costs of the five types of outpatient treatments ranged from $837 to $3334 per episode, and varied by both direct factors (e.g. hours of treatment, treatment retention) and indirect factors (e.g. cost of living, staff level, case-load variation)., Conclusions: These adolescent treatment cost estimates are examined in terms of their calculation, variability by condition, variability by site within condition and comparability with previous DATCAP results from outpatient drug-free programs for adults. Future research will integrate treatment outcomes and costs to complete cost-effectiveness and benefit-cost analyses of the five therapies.

Aims: This paper provides a description of the rationale, study design, treatments and assessment procedures used in the Cannabis Youth Treatment (CYT) experiment., Design: CYT was designed to (a) test the relative effectiveness, cost and benefit-cost of five promising treatment interventions under field conditions and (b) provide evidence based manual-guided models of these interventions to the treatment field., Setting: The study involved two community-based treatment programs and two major medical centers., Participants: Participants were 600 adolescents recruited from the regular intake who were between the ages of 12 and 18, had used marijuana in the past 90 days, and met one or more criteria of dependence or abuse., Interventions: Participants were randomly assigned to one of five interventions: Motivational Enhancement Therapy (MET), Cognitive Behavioral Therapy (CBT), Family Support Network (FSN), Adolescent Community Reinforcement Approach (ACRA), or Multidimensional Family Therapy (MDFT)., Measurements: Self-report data were collected at intake, 3, 6, 9 and 12 months post discharge using the Global Appraisal of Individual Needs (GAIN), as well as several supplemental self-reports, collateral reports, urine testing, and service logs., Findings: This paper reports on the study's implementation including the psychometric properties of the measures (alphas over 0.8), validity of self-report (kappa over 0.6), high rates of treatment completion (81% completed two or more months), and high rates of follow-up (over 94% per wave)., Conclusions: The feasibility of implementing the CYT manual-guided treatment and quality assurance model in community-based adolescent treatment programs is discussed.

Aims: Risk factors among adolescent substance abusers have been shown to correlate with substance use severity. Characteristics related to severity, such as demographic and family factors, peer influences, psychiatric co-morbidity and HIV risk behaviors, are examined for a sample of adolescent cannabis users entering treatment., Design: These data are from a clinical trial study utilizing blocked random assignment of clients to one of five treatment conditions. The study targeted adolescents entering outpatient treatment for primarily cannabis abuse or dependence., Setting: Treatment and research facilities in four metropolitan areas of the US were used to recruit study participants. Treatment was delivered in outpatient drug-free settings., Participants: Participants were 600 clients, ages 12-18, admitted to outpatient substance abuse treatment programs for cannabis problems, 96% with DSM-IV diagnoses of substance abuse or dependence, with the remaining 4% having at least one symptom of dependence plus significant problems indicating need for treatment., Measurements: The Global Appraisal of Individual Needs (GAIN) was used to collect the information presented in this paper. The GAIN incorporates DSM-IV criteria for substance use disorders, conduct disorder and attention deficit hyperactivity disorder, as well as dimensional (scale) measures for physical and mental health., Findings: All participants reported at least one symptom of substance use disorders, and 46% met the DSM-IV criteria for substance dependence, while 50% met criteria for a diagnosis of abuse. Only 20% of the participants perceived any need for help with problems associated with their drug or alcohol use. Clients participating in the study typically presented multiple problems at treatment entry, most often including conduct disorder, attention deficit hyperactivity disorder (ADHD), internal (mental) distress, and physical health distress. The co-occurrence of conduct disorder and ADHD was found in 30% of the sample. Clients meeting criteria for substance dependence tended to have more co-occurring problems and significantly less denial at admission., Conclusions: The characteristics of this sample exemplify the complex nature of adolescent substance use and abuse among adolescents entering outpatient treatment programs. Patterns of co-occurring problems are at rates comparable to those found in other clinical studies. Those with more severe substance use disorders tend to manifest more problems of social functioning, more mental health problems, and physical health problems. Implications of these findings are discussed in terms of treatment needs, challenges, and prognostic implications.

The ability of human neutrophils to aid in defense against pulmonary infection with Mycobacterium tuberculosis is controversial. In this study, we have shown that neutrophils respond to and phagocytose M. tuberculosis in human lesions. Neutrophils from healthy individuals were able to kill significant fractions of an inoculum of M. tuberculosis within 1 h of phagocytosis, and this ability was enhanced by tumor necrosis factor alpha but not by gamma interferon. The mycobactericidal mechanism was nonoxidative, as inhibitors of reactive oxygen or reactive nitrogen intermediates did not interfere with killing. However, the mycobactericidal mechanism was associated with increased exposure of intracellular M. tuberculosis to neutrophil defensins. In vitro, human neutrophil peptides 1 to 3 were not able to kill the bacilli even at much higher levels. These studies support the concept that human neutrophils are directly involved in defense against infection with M. tuberculosis.