Patterns of substance use among adolescents: A systematic review.

PURPOSE: This review characterizes empirically derived patterns of multiple (multi-) substance use among adolescents. A secondary objective was to examine the extent to which mental health symptomatology was included in the empirical analyses examining substance use patterns. METHODS: Eligible studies included those that used cluster-based approaches, included the assessment of at least two different substances, and were based on study samples with mean ages between 11 and 18 years. 4665 records were screened including 461 studies for full-text screening. RESULTS: 70 studies were included with common clusters being: low use, single or dual substance use, moderate general multi-use, and high multi-use. The most common patterns of single or multi-substance use were: alcohol only, alcohol with cannabis and/or tobacco, and use of alcohol, tobacco, and cannabis with and without other drugs. Lower socioeconomic status, older age, and male gender were consistent predictors of multi-use clusters. Only 37 % of studies compared differences in levels of mental health across clusters with symptoms consistently associated with a greater likelihood of multi-use. Only 29 % of studies included mental health indicators in cluster-based analyses, with over half identifying distinct mental health and substance use clusters. Fit indices in cluster analyses and measurement properties of substance use were heterogeneous and inconsistently reported across studies. CONCLUSIONS: Distinct patterns of substance use were derived but methodological differences prevented direct comparison and reduced capacity to generalize across studies. There is a need to establish standardized methodological approaches to identify robust patterns of substance use to enhance etiological, prognostic, and intervention research.

Development of a polymerase chain reaction (PCR) test for the detection of virulent forms of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a marine bacterium and some strains cause gastroenteritis in humans. Clinical isolates are thought to possess virulence factors that are absent from the majority of environmental isolates. Use of randomly amplified polymorphic DNA (RAPD)-PCR produced a unique 600 bp amplicon (band Y) in the majority of clinical isolates and rarely in environmental isolates tested. The DNA from band Y was cloned and sequenced and found to code for an outer membrane protein (OMP). Two polymerase chain reaction (PCR) primers were designed to specifically amplify a 200 bp unique sequence from presumptive virulent strains (PCR-OMP). The virulence of 23 clinical and 32 environmental isolates was assessed in cytotoxicity tests by treatment of Caco-2 cells with extracellular products (ECPs). All but two of the clinical isolates (91%) were positive for the 200 bp PCR-OMP and their ECPs produced a significantly higher (p < 0.05) lactate dehydrogenase (LDH) release (mean 72.88%) than the ECPs of environmental isolates (mean 15.3%) with the exception of one environmental isolate that produced the 200 bp amplicon. A positive 200 bp PCR-OMP is strongly correlated with virulence, as determined by the cytotoxicity assay, and identified virulent forms better than current PCR tests for tdh, trh or T3SS2.

Evidence for phosphonate usage in the coral holobiont.

Phosphonates are characterized by a stable carbon-phosphorus bond and commonly occur as lipid conjugates in invertebrate cell membranes. Phosphonoacetate hydrolase encoded by the phnA gene, catalyses the cleavage of phosphonoacetate to acetate and phosphate. In this study, we demonstrate the unusually high phnA diversity in coral-associated bacteria. The holobiont of eight coral species tested positive when screened for phnA using degenerate primers. In two soft coral species, Sinularia and Discosoma, sequencing of the phnA gene showed 13 distinct groups on the basis of 90% sequence identity across 100% of the sequence. A total of 16 bacterial taxa capable of using phosphonoacetate as the sole carbon and phosphorus source were isolated; 8 of which had a phnA+ genotype. This study enhances our understanding of the wide taxonomic and environmental distribution of phnA, and highlights the importance of phosphonates in marine ecosystems.

Phylogeny and molecular identification of vibrios on the basis of multilocus sequence analysis.

We analyzed the usefulness of rpoA, recA, and pyrH gene sequences for the identification of vibrios. We sequenced fragments of these loci from a collection of 208 representative strains, including 192 well-documented Vibrionaceae strains and 16 presumptive Vibrio isolates associated with coral bleaching. In order to determine the intraspecies variation among the three loci, we included several representative strains per species. The phylogenetic trees constructed with the different genetic loci were roughly in agreement with former polyphasic taxonomic studies, including the 16S rRNA-based phylogeny of vibrios. The families Vibrionaceae, Photobacteriaceae, Enterovibrionaceae, and Salinivibrionaceae were all differentiated on the basis of each genetic locus. Each species clearly formed separated clusters with at least 98, 94, and 94% rpoA, recA, and pyrH gene sequence similarity, respectively. The genus Vibrio was heterogeneous and polyphyletic, with Vibrio fischeri, V. logei, and V. wodanis grouping closer to the Photobacterium genus. V. halioticoli-, V. harveyi-, V. splendidus-, and V. tubiashii-related species formed groups within the genus Vibrio. Overall, the three genetic loci were more discriminatory among species than were 16S rRNA sequences. In some cases, e.g., within the V. splendidus and V. tubiashii group, rpoA gene sequences were slightly less discriminatory than recA and pyrH sequences. In these cases, the combination of several loci will yield the most robust identification. We can conclude that strains of the same species will have at least 98, 94, and 94% rpoA, recA, and pyrH gene sequence similarity, respectively.

Photobacterium rosenbergii sp. nov. and Enterovibrio coralii sp. nov., vibrios associated with coral bleaching.

Six new Vibrio-like isolates originating from different species of bleached and healthy corals around Magnetic Island (Australia) were investigated using a polyphasic approach. Phylogenetic analyses based on 16S rRNA, recA and rpoA gene sequences split the isolates in two new groups. Strains LMG 22223(T), LMG 22224, LMG 22225, LMG 22226 and LMG 22227 were phylogenetic neighbours of Photobacterium leiognathi LMG 4228(T) (95.6 % 16S rRNA gene sequence similarity), whereas strain LMG 22228(T) was related to Enterovibrio norvegicus LMG 19839(T) (95.5 % 16S rRNA gene sequence similarity). The two new groups can be distinguished from closely related species on the basis of several phenotypic features, including fermentation of d-mannitol, melibiose and sucrose, and utilization of different compounds as carbon sources, arginine dihydrolase activity, nitrate reduction, resistance to the vibriostatic agent O/129 and the presence of fatty acids 15 : 0 iso and 17 : 0 iso. The names Photobacterium rosenbergii sp. nov. (type strain LMG 22223(T)=CBMAI 622(T)=CC1(T)) and Enterovibrio coralii sp. nov. (type strain LMG 22228(T)=CBMAI 623(T)=CC17(T)) are proposed to accommodate these new isolates. The G+C contents of the DNA of the two type strains are respectively 47.6 and 48.2 mol%.

A survey for selected viral, chlamydial, and parasitic diseases in wild dusky-headed parakeets (Aratinga weddellii) and tui parakeets (Brotogeris sanctithomae) in Peru.

Thirty-eight free-ranging dusky-headed parakeets (Aratinga weddellii) and 13 tui parakeets (Brotogeris sanctithomae) were caught and released in Parque Nacional del Manu in southeastern Peru from 19 July to 5 August 1993. Blood and fecal samples were collected and sera were evaluated for titers to Pacheco's disease herpesvirus, psittacine polyomavirus, paramyxovirus-1, and Chlamydia psittaci. Fecal samples were examined for evidence of ascarid or coccidial infection by fecal flotation, and blood smears were examined for hemoparasites. Five (50%) of 10 A. weddellii serum samples tested by complement fixation (CF) for psittacine polyomavirus antibodies were positive, and three (19%) of 16 A. weddellii samples tested by virus neutralization (VN) for psittacine polyomavirus antibodies were positive, yielding a total of 8 (38%) of the 21 A. weddellii samples positive for psittacine polyomavirus. Based on CF for herpesvirus, four (11%) of 38 A. weddellii samples had antibodies against herpesvirus. All B. sanctithomae were negative for psittacine polyomavirus and psittacine herpesvirus. Thirty-five of the A. weddellii tested were negative for Chlamydia psittaci by CF, latex agglutination, and elementary body agglutination, and all B. sanctithomae were negative for Chlamydia psittaci by the CF test. Nine A. weddellii and eight B. sanctithomae evaluated for paramyxovirus-1 titers by the hemagglutination inhibition test were negative. All fecal samples were negative for ascarids or coccidia by fecal flotation, and all blood smears were negative for hemoparasites by direct microscopic examination. This is the first known description of psittacine polyomavirus and psittacine herpesvirus in free-ranging parrots. Serologic evidence of Pacheco's disease herpesvirus in wild A. weddellii is interesting in light of the fact that Aratinga spp. are considered to be possible carriers of this virus in captivity.

Transepidermal induction of contact hypersensitivity in mice with a water-soluble hapten.

An experimental analysis has been conducted on the capacity of a highly-reactive, water soluble hapten, trinitrobenzene sulfonic acid, to induce contact hypersensitivity when applied epicutaneously to body wall skin of normal mice. Application of plastic chambers containing the hapten to murine skin for as little as 1 h produced readily detectable sensitization in several genetically disparate inbred strains. Moreover, the efficiency of sensitization was found to be similar to that following epicutaneous application of this hapten's lipid-soluble cogener, trinitrochlorobenzene. Using this approach, it has been determined that UVB radiation, intradermally injected tumor necrosis factor-alpha, and epicutaneously applied cis-urocanic acid can impair contact hypersensitivity induction by transepidermally delivered trinitrobenzene sulfonic acid, and that animals that fail to become sensitized proceed to acquire hapten-specific unresponsiveness. It is concluded that epicutaneous sensitization to chemically reactive, water-soluble molecules is experimentally attainable if precautions are taken to insure that contact between the hapten solution and the cutaneous surface is maintained for at least 1 h. Understanding the cellular and molecular mechanisms of sensitization and tolerance induction by epicutaneously applied water soluble haptens may prove to be important in understanding the pathogenesis of allergic contact dermatitis that develops to chemicals in the industrial setting, in the environment, and in the clinic.

Culture media for the differentiation of isolates of Yersinia ruckeri, based on detection of a virulence factor.

Strains of the bacterial fish pathogen Yersinia ruckeri were identified with the API 20E system and distinguished on the basis of whole cell agglutination with antisera, sorbitol fermentation and polymyxin B sensitivity. Strains which were shown to possess the virulence-associated heat-sensitive factor (HSF) were shown to grow preferentially on culture media containing sodium dodecyl sulphate (SDS) and to produce a creamy deposit around the colonies. By contrast, strains lacking this factor (HSF-) grew poorly and without forming a deposit. Enhancement of the differentiation between the two types was shown by the incorporation of Coomassie brilliant blue dye into agar containing 1% SDS, and the uptake of Coomassie blue and Congo red was shown to be temperature-dependent. Most strains tested were shown to belong to serotype I, and were sensitive to polymyxin and did not ferment sorbitol. With the medium developed most serotype I strains but not those of other serotypes were shown to possess HSF. It is suggested that the medium is used in epidemiological studies of Y. ruckeri.

Internalization of Ia molecules into Birbeck granule-like structures in murine dendritic cells.

Dendritic cells isolated from the draining lymph nodes of mice sensitized epicutaneously with hapten are potent antigen-presenting cells and contain Birbeck granules and cored tubules characteristic of antigen-activated epidermal Langerhans cells. We used immunogold labeling and transmission electron microscopy to follow the internalization of Ia molecules in these antigen-presenting cells. We found that Ia molecules were internalized into Birbeck granule-like structures in the antigen-activated dendritic cells. Computer reconstruction of serial sections of the dendritic cells demonstrated that these structures span the cytoplasm from the cell membrane to the nuclear membrane and are associated with lysosomes. The internalization of Ia molecules into these structures supports the hypothesis that the Birbeck granule-like structures are derived from the cell membrane and are involved in the antigen-processing/presenting function of the dendritic cells.

Assessment of X bends in patients with atypical X chromosome phenotypes.

Several patients with X chromosome structural abnormalities have been more severely affected clinically than expected. Since bends at Xq13-21 have been associated with inactivation, the authors scored bends retrospectively in 62 patients with X chromosome aneuploidy and 21 cases with structural abnormalities of the X chromosome. They found that patients with 2 X inactivation sites where one X was structurally abnormal had significantly fewer cells with X bends than normal 46,XX. In addition, these patients also showed X bends on the normal X more often than would be expected if non-random X inactivation of the abnormal X chromosome was occurring. Five of the 6 patients with a short or long arm deletion or paracentric inversion of Xq were mentally retarded or had other congenital anomalies not usually associated with Turner syndrome. This suggests to them that these clinical findings may be related to interference with X inactivation patterns in cells with a structurally abnormal X chromosome.

Evidence that cutaneous antigen-presenting cells migrate to regional lymph nodes during contact sensitization.

These studies address the hypothesis that Ag-bearing epidermal Langerhans cells migrate to the regional lymph node during contact sensitization and function as APC. Skin from C3H mice was grafted onto BALB/c nude mice, and 7 or 14 days later, the recipients were sensitized with FITC through the grafts. APC from lymph nodes draining the site of sensitization were capable of sensitizing C3H recipients to FITC. Because sensitization is MHC restricted, only cells reaching the lymph node from the grafted skin could have induced contact hypersensitivity in C3H mice. Examination of the FITC+ draining lymph node cells by immunofluorescence and immunoelectron microscopy demonstrated that all were Ia+, most were F4/80+, and some contained Birbeck granules. These studies demonstrate that Ia+, FITC+ cells from the skin, at least some of which are Langerhans cells, leave the skin after epicutaneous sensitization with FITC and participate in the initiation of the contact hypersensitivity response within the regional lymph node.

The cloning and expression of a gene encoding haemolytic activity from the fish pathogen Renibacterium salmoninarum.

A gene encoding haemolytic activity from Renibacterium salmoninarum (strain PPD) was cloned into Escherichia coli using the cosmid vector pHC79, and subsequently subcloned on a 1.6 kbp SAlI fragment into pBR328. Southern blot hybridisation revealed that a homologous sequence is found in other strains of R. salmoninarum.

Virulence of Yersinia ruckeri serotype I strains is associated with a heat sensitive factor (HSF) in cell extracts.

Cell extracts of Yersinia ruckeri (serotype I) were examined by SDS-PAGE and Western blotting. An unusual band, termed heat-sensitive factor (HSF) was observed in extracts of virulent strains only. It is thought to be lipid in nature; no differences could be detected in the region of the band in protein profiles of virulent and avirulent strains. When trout were infected either by intraperitoneal injection or bath immersion, mortalities occurred only with HSF+ strains. The HSF appears to be an important virulence determinant of Y. ruckeri.

Malignant mesothelioma of tunica vaginalis testis.

Malignant mesothelioma of the tunica vaginalis is rare, but sometimes curable. It is similar to malignant mesothelioma of the peritoneum and of the pleura, and is likewise associated with asbestos exposure. We report a case, with correlative computed tomography, ultrasound, and gross pathology images that demonstrate tiny tumor implants studding the vaginalis testis. The literature is reviewed.

c-myc oncoprotein levels in bladder cancer.

The protein coded by the oncogene c-myc, p62c-myc, was measured using monoclonal antibodies and flow cytometry in nuclei derived from paraffin-wax sections of transitional cell carcinomas of the human bladder. Superficial disease (stages pTa and pT1) which did not recur within 5 years of diagnosis had significantly higher oncoprotein levels than those which did recur or were muscle-invasive (stage pT2 or greater) at presentation (P less than 0.01). These preliminary findings indicate that oncoprotein levels might have prognostic significance for bladder cancer.

Oncogene expression in ovarian cancer: a pilot study of c-myc oncoprotein in serous papillary ovarian cancer.

The nuclear-associated protein product of the c-myc gene, p62c-myc, was assayed simultaneously with total DNA using flow cytometry in nuclei extracted from archival biopsies of serous papillary carcinoma of the ovary. The oncoprotein was probed with a synthetic peptide-induced mouse monoclonal antibody which was subsequently labeled with a fluorescent rabbit anti-mouse immunoglobulin and DNA was assayed using the nucleic acid fluorochrome propidium iodide. Serous papillary ovarian carcinoma expressed significantly higher p62c-myc levels compared with normal ovary (P less than 0.00003 Mann-Whitney U test). Biopsies classified as "borderline" low-potential malignancy exhibited levels between normal ovary and carcinoma. The difference between normal and "borderline" was significant at P less than 0.003, but no difference between "borderline" and frankly invasive biopsies was observed, P = 0.149. There was no difference among the histological grades of carcinomas. All normal ovaries had diploid DNA content as did 5/6 cases of "borderline" malignancy. The majority of cases of carcinoma, 28/36, were aneuploid. There was a statistically significant difference in the distribution of aneuploidy, P less than 0.005, between invasive carcinomas and those classified as "borderline" low-potential malignancy.

Comparison of the immunosuppressive effects of cyclosporine, lipid-soluble anesthetics, and calmodulin antagonists. Response to exogenous interleukin 2.

The purpose of these investigations was to compare the immunosuppressive mechanism of cyclosporine (CsA) with those of lipid-soluble local anesthetics and calmodulin antagonists. Chlorpromazine (CPZ) and pentobarbital (PB) both inhibit lymphocyte activation by attenuating sodium and potassium ion potentials. CPZ and N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W-7) can also block calcium-dependent activation processes by inhibition of calmodulin and protein kinase C. All four compounds were found to suppress human and murine lymphoproliferation to both alloantigen or mitogen in a dose-dependent and saturable manner. Exogenous interleukin-2 (IL-2) restored mitogenic responsiveness to cultures suppressed using W-7 and CsA, but not to lymphocytes suppressed with either CPZ or PB. Cytofluorographic analysis revealed that the degree of suppression in drug-treated lymphocytes was significantly correlated with the surface expression of receptors for transferrin and interleukin-2. Inhibition of IL-2 activation by PB was demonstrated to result from a blockade of the mitogenic growth factor signal using the IL-2-dependent cell line HT-2. Thus, the mechanism of action of cyclosporine can be differentiated from those of anesthetic immunosuppressants at the level of responsiveness to interleukin-2. The data support the hypothesis that cyclosporine may be an antagonist of calmodulin that selectively blocks early events in T lymphocyte activation leading to IL-2 synthesis, but does not inhibit the expression or function of the IL-2 receptor.

Pleural lipoma. Diagnosis by computed tomography.

Until recently, a definitive diagnosis of lipoma in the thorax could only be established by thoracotomy. We undertook this study to determine if chest CT could provide such an answer. Among 4,000 chest CT scans, six patients were found to have lipoma according to the following selected criteria: CT features of a pleural mass; a lesion showing completely homogeneous density with CT numbers indicating fat, and exclusion of other fatty lesions. In these six patients, the lipoma was an incidental finding, four were men, the mean age was 64.3 years, one-half were obese, and none had chest pains or dyspnea. Lesions varied in size from 2 to 4 cm and occurred along the chest wall. The CT numbers of the masses ranged from -54 to -129. None developed malignancy. In conclusion, we recommend clinical and chest CT follow-up for the asymptomatic patient who fulfills our CT criteria for lipoma. Biopsy or resection is recommended for lesions that are inhomogeneous.