A Strengths-Based Longitudinal Career Intervention for Junior Secondary School Students with Special Educational Needs: a Mixed-method Evaluation.

The mixed-method study reported here was designed to evaluate a strengths-based career intervention program for secondary school students with mild special educational needs (SEN). A sample of 32 SEN students (19 boys: 13 girls) from 5 inclusive schools in Hong Kong were recruited to a treatment group. An additional 32 SEN students (19 boys: 13 girls) were selected to form the control group matched for age, gender and parents' education level. The special needs exhibited by both groups were in areas of literacy and numeracy, attention deficits, and social-emotional problems, but did not include severe or complex disabilities. Participants in both groups responded to pre- and post-intervention questionnaires covering career development self-efficacy, personal and social development self-efficacy, and meaning in life. As a follow-up, two teachers and three social workers providing support to SEN students, and the 32 participants were interviewed several months after the intervention. Interviews also took place with teachers, social workers and students to evaluate the perceived effects of the intervention. Findings indicated significant interactions between Time 1 and Time 2, and between groups (control vs. treatment) in personal goal-setting, career goal-setting, and the presence of meaning in life. Additionally, several themes were identified from the interviews suggesting that the intervention did have positive effects on SEN students' career, personal and social development self-efficacy, and acquisition of meaning in life.

Changes of serum cytokine profiles in Korean vivax malarial patients after chemotherapy.

Changes of serum cytokines including interferon gamma inducible protein-10 (IP-10) chemokine were analyzed in a total of eight vivax malarial patients before and after chemotherapy by immunostaining for human cytokine antibodies attached to membranes. IP-10 was strongly reactive in the period before chloroquine/primaquine combined chemotherapy and disappeared after chemotherapy in all patients. Therefore, IP-10 chemokine may be of importance in the pathogenesis of vivax malaria and a good marker of a complete cure.

Pharmacokinetics and metabolism of acriflavine in rats following intravenous or intramuscular administration of AG60, a mixture of acriflavine and guanosine, a potential antitumour agent.

Acriflavine (ACF; CAS 8063-24-9), a mixture of trypaflavine (TRF) and proflavine (PRF) at a ratio of 2:1 is being investigated in rodents as an anticancer agent. However, its pharmacokinetics have not been investigated in mammals. Guanosine is known to potentiate the anticancer activity of some compounds. The pharmacokinetics of AG60, a 1:1 mixture of ACF and guanosine, were therefore investigated in rats. Rats were given 2 or 10 mg kg(-1) AG60 by intravenous bolus or 6 or 30 mg kg (-1) intramuscularly. An HPLC-based method was developed to analyse the levels of TRF, PRF, and their metabolites in plasma, bile, urine and tissue homogenates. The plasma concentrations of TRF and PRF decreased rapidly after intravenous administration and more slowly after intramuscular administration. Both TRF and PRF were distributed widely, most notably in the kidney, and were eliminated slowly. Three glucuronosyl conjugate metabolite peaks were tentatively identified in the bile. The intramuscular route leads to a prolongation of TRF or PRF plasma levels, and the systemic exposures for both TRF and PRF were both relatively high. These observations indicate that the intramuscular route may be the best way to administer AG60 for various clinical applications.

Early cysteine protease activity in excretory bladder triggers metacercaria excystment of Paragonimus westermani.

The cysteine proteases of Paragonimus westermani metacercaria are known to initiate metacercaria excystment. However, their secretory sites, such as the intestine, tegument, and excretory bladder are not well defined. In this study, the metacercariae were labeled with bromodeoxyuridine (BrdU) to distinguish the initial activation sites. BrdU was labeled mainly at the excretory bladder and the excretory granules of the metacercariae and the newly excysted juvenile worms. This result shows that early 'defecation' of the proteases from the excretory bladder may accelerate the excystment of P. westermani metacercariae.

A cathepsin F of adult Clonorchis sinensis and its phylogenetic conservation in trematodes.

A novel 28 kDa cysteine protease (Cs28CF) secreted by the hepatobiliary trematode, Clonorchis sinensis was identified. The protease was purified from the excretory-secretory products (ESP) of the adult worm using DEAE-ion exchange and Arginine-Sepharose 4B chromatography. It showed a high activity between pH 6.5 and 7.5 in a dithiothreitol (DTT)-dependent manner. Inhibitors specific to cysteine proteases down-regulated the activity. Addition of Cs28CF to monkey cholangiocyte cultures resulted in approximately 95% cell death after 7 days. The full-length cDNA (1078 bp) encoded a single peptide of 328 amino acids (aa) with an N-terminal hydrophobic sequence, an ERFNAQ motif in the propeptide and a mature domain. Expression of mRNA transcripts of Cs28CF was observed in both the metacercaria and adult stages. Bacterially expressed recombinant protein exhibited a specific antibody reaction with clonorchiasis sera. Deduced aa exhibited 52-76% sequence identity with the cathepsin F analogues from other organisms. A novel E/DXGTA motif was recognized in the propeptide region. Phylogenetic analysis of 63 papain family members revealed that the trematode cysteine proteases formed 2 major clades of cathepsins F and L. The trematode cysteine proteases classified as cathepsin F shared higher homology among themselves than those classified as cathepsin L. Cathepsin F is phylogenetically conserved in the trematode parasites as well as in mammals.

Purification and localization of a 10 kDa calcareous corpuscle binding protein of Spirometra mansoni plerocercoid.

In cestode parasites, calcareous corpuscles are thought to be associated with a number of intracellular physiologies by regulating the trafficking of mineral components. We previously separated these particular components by Ficoll, and their binding proteins of 10 kDa and 35 kDa in the Spirometra mansoni plerocercoid (sparganum). In the present study, we purified a 10 kDa protein employing octyl-Sepharose CL-4B affinity chromatography. Anti-serum raised against the purified protein showed specific reactions to the calcareous corpuscles of the worm section by immunohistochemistry, and recognized the 10 kDa protein by immunoblotting.

Determination of recombinant human epidermal growth factor (rhEGF) in a pharmaceutical formulation by high performance liquid chromatography with electrochemical detection.

A novel HPLC method with electrochemical detection has been developed for the determination of recombinant human epidermal growth factor (rhEGF) in pharmaceutical products. rhEGF was separated from other components in formulation on a reversed-phase C18 column with 24% acetonitrile in 0.1 M phosphate buffer (pH 4.75). The optimum electrochemical oxidation of EGF was obtained at 0.85 V vs. Ag/AgCl in a glassy carbon working electrode due to electroactive tyrosine, tryptophan, methionine, and arginine residues. The quantitation range was from 1.0 to 200 ng of rhEGF with the linear correlation coefficient greater than 0.999. The method was successfully applied for the quantitation of rhEGF in a pharmaceutical preparation.

The Lesbian Internalized Homophobia Scale: a rational/theoretical approach.

This article reports the development and psychometric properties of a new scale that measures internalized homophobia in lesbians: the Lesbian Internalized Homophobia Scale (LIHS). This 52-item measure was developed using a rational/theoretical approach of test construction and includes five subscales. Research findings, based on a sample of 303 female participants, supported the reliability and validity of the LIHS in assessing internalized homophobia in lesbians. Implications for research and practice are discussed.

Clonorchis sinensis: molecular cloning and characterization of 28-kDa glutathione S-transferase.

A 28-kDa glutathione S-transferase (Cs28GST) was purified from a Clonorchis sinensis cytosolic fraction through anion-exchange and glutathione-affinity column chromatographies. A monoclonal antibody raised against Cs28GST reacted specifically to the C. sinensis antigen among trematode proteins. A putative peptide of 212 amino residues deduced from a cDNA clone appeared homologous with 28-kDa GST of trematodes, and its secondary structural elements predicted a GSH-binding site. Recombinant Cs28GST showed GST enzyme activity with CDNB substrate and was sensitive to the model inhibitors. The recombinant Cs28GST was antigenically indistinguishable from the native form and was recognized specifically by C. sinensis-infected human sera. The Cs28GST was localized in the tegument and underlying mesenchymal tissues. It is suggested that Cs28GST may play significant physiological roles against bioreactive molecules and be a useful reagent for serodiagnosis of clonorchiasis.

Determination of recombinant human epidermal growth factor (rhEGF) in a pharmaceutical preparation by capillary electrophoresis.

A simple assay method of recombinant human epidermal growth factor (rhEGF) in a pharmaceutical preparation was studied and validated by capillary electrophoresis (CE) using micellar electrokinetic chromatography (MEKC) techniques. Factors affecting the migration behavior and separation performances of the peptide; type of buffer, pH, buffer concentration, and concentration of sodium dodecyl sulfates (SDS) were investigated to optimize the analytical performance. CE was performed using running buffer, 50.0 mM borate (pH 8.5) containing 12.5 mM SDS at 20 kV of the applied voltage. Calibration curves for the rhEGF showed good linearity (r>0.999) over the wide dynamic range from 1.25 to 100 microg/ml. Sample analysis was performed by using standard addition method to eliminate the matrix effects of dosage vehicle. This method is assumed to be useful for quality control (QC) of various forms of pharmaceutical products of the peptide.

Structural and immunological characteristics of a 28-kilodalton cruzipain-like cysteine protease of Paragonimus westermani expressed in the definitive host stage.

A complete cDNA sequence encoding a 28-kDa cruzipain-like cysteine protease of adult Paragonimus westermani, termed Pw28CCP, was isolated from an adult cDNA library. The cDNA contained a single open reading frame of 975 bp encoding 325 amino acids, which exhibited the structural motif and domain organization characteristic of cysteine proteases of non-cathepsin Bs including a hydrophobic signal sequence, an ERFNIN motif, and essential cysteine residues as well as active sites in the mature catalytic region. Analysis of its phylogenetic position revealed that this novel enzyme belonged to the cruzipain-like cysteine proteases. The sequence of the first 13 amino acids predicted from the mature domain of Pw28CCP was in accord with that determined from the native 28-kDa enzyme purified from the adult worm. Expression of Pw28CCP was observed specifically in juvenile and adult worms, with a location in the intestinal epithelium, suggesting that this enzyme could be secreted and involved in nutrient uptake and immune modulation. The recombinant protein expressed in Escherichia coli was used to assess antigenicity by immunoblotting with sera from patients with active paragonimiasis and from those with other parasitic infections. The resulting sensitivity of 86.2% (56 of 65 samples) and specificity of 98% (147 of 150 samples) suggest its potential as an antigen for use in immunodiagnosis.

IgG antibody responses in early experimental sparganosis and IgG subclass responses in human sparganosis.

Antigenic components in the crude extracts of Spirometra mansoni plerocercoid were analyzed in early experimental infections and in IgG subclass observed in clinical sparganosis. By IgG immunoblot, sera obtained serially from experimental mice, fed 5 spargana each, were reacted with the crude extracts. Protein bands at 36-26 kDa and 103 kDa showed positive reactions since two weeks after infection. In a differential immunoblot, in which a monospecific antibody against sparganum chymase at 36 kDa was pre-treated, the reactions at 36-26 kDa disappeared, indicating that the sparganum chymase and its degradation products invoked IgG antibody reactions. When 69 patients sera of human sparganosis were examined for their IgG subclass responses, IgG4 levels showed the highest reaction which was followed by IgG1. The IgG4 antibody also reacted mainly with 36-31 kDa protease. These results indicate that 36 kDa chymase of S. mansoni plerocercoid is the main antigenic component inducing IgG antibody response in early stage of experimental sparganosis and for specific IgG subclass reactions in human sparganosis.

Purification of a 68-kDa cysteine proteinase from crude extract of Pneumocystis carinii.

The present study intended to verify activities of cysteine proteinase of Pneumocystis carinii from rats and to purify the enzyme. In order to exclude the contamination of host-derived enzymes, concentrates of P. carinii was primarily treated with a mixture of proteinase inhibitors before lysis of P. carinii. A 68-kDa cysteine proteinase was finally purified from the crude extract of P. carinii by 4 sequential chromatographic methods. The enzyme showed an optimal activity at pH 5.5 in 0.1 M sodium acetate, and its activity was specifically inhibited by L-trans-epoxy-succinylleucylamido (4-guanidino) butane (E-64) and iodoacetic acid, suggesting that the enzyme is a cysteine proteinase. The 68-kDa proteinase weakly digested macromolecules such as collagen, hemoglobin and fibronectin. The present study demonstrated the activity of cysteine proteinase at the 68-kDa band of P. carinii, and purified and characterized the molecule.

Differential expression of the 27 kDa cathepsin L-like cysteine protease in developmental stages of Spirometra erinacei.

The 27 kDa cathepsin L-like cysteine protease of Spirometra erinacei plerocercoid is known to play an important function in tissue penetration, nutrient uptake and immune modulation in human sparganosis. In the present study, the expression of this enzyme was examined at different developmental stages of S. erinacei including immature egg, coracidium, plerocercoid in tadpole and rat, and adult. Proteolytic activity against carboxybenzoyl-phenylalanyl-arginyl-7-amino-4-methylcoumarin was detected in the extracts of coracidia and plerocercoid while no activity was observed in those of immature egg and adult. The specific activity in coracidial extracts was lower than that in the plerocercoid. Reverse transcription-polymerase chain reaction and Northern blot analysis demonstrated that the gene was expressed in the coracidium and plerocercoid but not in immature egg and adult. These results suggest that the 27 kDa cysteine protease is only expressed in the stages involving active migration of the parasite in the host tissue.

Persisting antibody reaction in paragonimiasis after praziquantel treatment is elicited mainly by egg antigens.

Antibody responses in serum and cerebrospinal fluid (CSF) samples from patients with active and chronic paragonimiasis and in sera from patients on whom follow-up studies were done after praziquantel treatment were analyzed using antigens of Paragonimus westermani prepared from eggs, metacercariae, juveniles of 4- and 7-week old, adult worms and recombinant protein of 28 kDa cruzipain-like cysteine protease (rPw28CCP). The patient sera/CSFs of active and chronic paragonimiasis revealed strong antibody reactions against the crude extracts of 4- and 7-week old juveniles as well as against those from egg and adult. rPw28CCP also showed specific reaction to the sera with active paragonimiasis. After the treatment, levels of specific antibodies in the sera gradually decreased to negative range in most patients. In some cases with persisting high antibody levels, however, the reactions at 27 kDa egg protein were sustained throughout the observation period of 34 months. The reactions at 35 and 32 kDa in adult extract and rPw28CCP disappeared rapidly after the treatment. Persistent antibody reactions even after successful treatment are provoked by continuous antigenic challenge from eggs which were not resolved by treatment.

Partial characterization of a 17 kDa protein of Clonorchis sinensis.

A 17 kDa protein from Clonorchis sinensis adults was purified by a procedure including Sephacryl S-200 HR gel filtration and Q-Sepharose anion exchange chromatography. The protein was proved to be a cysteine protease as it showed hydrolytic activity toward Cbz-Phe-Arg-AMC in the presence of dithiothreitol and was inhibited by specific inhibitors such as iodoacetic acid or trans epoxy-succinly-L-leucyl-amido(4-guanidino) butane. The polyclonal antibody raised against the protein reacted to 17 kDa proteins of trematodes such as Paragonimus westermani, Fasciola hepatica, Opisthorchis viverrini, Gymnophalloides seoi, and Metagonimus yokogawai. The antibody recognized the 17 kDa and 16 kDa cysteine proteases purified from C. sinensis, P. westermani, and G. seoi as well. These results suggest that the 17 kDa protein may be a cysteine protease commonly present in trematodes.

Seroprevalence of toxoplasmosis in the residents of Cheju island, Korea.

This study was performed to evaluate the epidemiological status of toxoplasmosis among the residents of Cheju island. The sera of local students from 18 high schools (boys 2110, girls 2460) and those of adults (474 admitted to Cheju Chungang General Hospital) were collected and checked for the IgG antibody titers against Toxoplasma gondii. Serum samples collected from both the students and adults showed sero-positive rate of 5.5% and 12.9%, respectively. Although the rates were not significantly different between the sexes (5.4% for the boys and 5.5% for the girls attending school), the geographical difference showed a significant difference between the urban (4.6-6.9%) and rural areas (5.6-8.8%) (p < 0.05). Based on the high positive rates, it should be necessary to control toxoplasmosis in Cheju island.

Paragonimus westermani: a cytosolic glutathione S-transferase of a sigma-class in adult stage.

We purified cytosolic glutathione S-transferase (GST) of adult Paragonimus westermani monitoring its activity with 1-chloro-2,4-dinitrobenzene (CDNB). The enzyme was purified 18.4-fold to electrophoretic homogeneity with 21% recovery rate through a three-step procedure. The purified enzyme (Pw28GST) has a subunit molecular weight of 28 kDa with an isoelectric point at 4.6. Monoclonal antibody (anti-Pw28GST) against Pw28GST did not cross-react with GSTs from other helminths. cDNA library was constructed in lambdaZAP II bacteriophage and screened with anti-Pw28GST. The corresponding gene containing a single open reading frame of 804 bp encoded 211 amino acids. The predicted amino acid sequence exhibited a higher homology with catalytic domain near N-terminus of class sigma GSTs (58%) than with schistosome 28-kDa GSTs (45-41%) or with class sigma GSTs themselves (33-31%). The sequence contained both Tyr-6 and Tyr-10 that are highly conserved in mammalian and helminth GSTs. The apparent K(m) value of a recombinant enzyme was 0.78 mM. Both native and recombinant enzymes showed the highest activity against CDNB, relatively weak activity against ethacrynic acid and reactive carbonyls, and no activity against epoxy-3-(p-nitrophenoxy)-propane. The activities were inhibited by bromosulfophthalein, cibacron blue, and albendazole, but not by praziquantel. These findings indicate that adult P. westermani has a class sigma GST.

Survey of the Korean population for 9 short tandem repeat loci.

The short tandem repeats with repeat units ranging from two to several nucleotides became a powerful tool in the field of forensic identification and paternity determination as well as for research in human gene mapping. Allele and genotype frequencies for 9 short tandem repeats including HUMCSF1PO, HUMTH01, HUMPLA2A1, HUMF13A01, HUMCYAR04, HUMLIPOL, HUMHPRTB, HUMCD4, and HUMFABP were determined using PCR and subsequent analysis of the PCR products by denaturing polyacrylamide gel electrophoresis followed by silver-staining. DNA samples were obtained from about 100 Korean people and amplified in a thermocycler adopting glass capillaries rather than traditional tubes. We found that the bovine serum albumin was an essential additive for the capillary PCR, presumably to coat the inner surface of the capillary which may adsorb Taq DNA polymerase. The capillary thermocycler was very effective in reducing the cycling time such that most of the amplification reactions could be finished within 30 min albeit the PCR product was less than that for the tube systems. All loci except HUMHPRTB met the Hardy-Weinberg expectations according to the exact test. The cumulative power of discrimination (PD) was 0.9999998 and the power of exclusion (POE) for the paternity test was a little low, being 0.9873989.

Effects of a peer-mediated aerobic conditioning program on fitness levels of youth with mental retardation: two systematic replications.

The generality of a peer-mediated exercise program designed to enhance aerobic fitness of 17 children with moderate and severe cognitive disabilities was evaluated. Two systematic replications of the program were conducted. Participants' ages, disability levels, and school settings varied. Target participants were paired with peers without disabilities. These students encouraged their peers to maintain requisite levels of exercise intensity and monitored their heart rates. A between-group multiple-baseline design was used to evaluate program effects. Results replicated earlier findings. Aerobic fitness of participants, measured by exercise heart-rates, improved when the exercise program was introduced. Results for individual participants reflected more variability than combined group data. Implications for overcoming motivational barriers and obtaining valued outcomes correlated with the program are discussed.