To investigate the relationship between 12 candidate genes responsible for water regulation, sodium metabolism and membrane ion transport and essential hypertension (EH) in the Chinese. Linkage analysis of EH was performed in 95 Chinese nuclear families including 477 subjects using a technique of fluorescence-based gene scanning with 12 microsatellite markers. Markers were selected on the chromosomal regions covering 12 candidate genes responsible for regulating water and sodium metabolism and membrane ion transport. These candidate genes included sodium hydrogen exchanger 3, sodium hydrogen exchanger 5, chloride bicarbonate exchanger 3, sodium calcium exchanger 1, mineralocorticoid receptor, plasma membrane calcium ATPase 2, ATPase,Na/K transporting alpha, a-adducin, SA gene, kidney epithelial sodium channel-gamma, vasopressin receptor 1A, and 11beta-hydroxysteroid dehydrogenase type 2 genes. Two-point non-parametric linkage analysis (NPL), maximum LOD score analysis and transmission/disequilibrium test (TDT) were performed using the GENEHUNTER software package. The NPL analysis and LOD score suggested a significant linkage at D12S398 (Z = 2.08, p<0.05 and LOD score = 1.26, p<0.01, respectively). TDT indicated a significant disequilibrium of transmission at the locus chi2 = 9.00, p < 0.005). No significant linkages were found at the other loci tested (p>> 0.05 or LOD < -1). In conclusion, D12S398, a marker near the vasopressin receptor 1A gene (V1AR), showed a positive linkage with EH based on the results of three statistical methods (NPL, LOD score, and TDT). This region warrants further exploration.

We have previously described the synthesis of a cytotoxic polymeric conjugate of spermine (Poly-SPM) which is able to inhibit the transport of polyamines (spermine, spermidine, and putrescine) into normal and malignant cells. Recent studies examining the toxicity of Poly-SPM in parental and multidrug resistant (MDR) cancer cells have revealed a cross-resistance in the MDR variant Dx5 to the toxic effects of the conjugate in the MDR-positive cells. There were also differences in spermine and putrescine uptake rates between parental and MDR-positive with the MDR-positive cells having a lower Vmax and a higher Km. The ability of this Poly-SPM to reverse MDR was examined in MDR variants (Dx5 cells) of the human sarcoma cell line MES-SA. The cells express high levels of the mdr1 gene product, P-glycoprotein, and are 25-to 60-fold resistant to doxorubicin (DOX), etoposide (VP-16), vinblastine (VBL), and taxol (TAX). Cytotoxicity was measured by the MTT [3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Poly-SPM (50 microM) lowered the drug concentration IC50 values in the Dx5 cells by 37-fold with VBL, 42-fold with DOX, 29-fold with VP-16, and 25-fold with TAX when compared to the control IC50 values without Poly-SPM. This reversal of resistance was concentration dependent, decreasing 17-fold with DOX, 6.1-fold with VBL, 19-fold with VP-16, and 5-fold with TAX when 25 microM Poly-SPM was used. No modulation was observed in the parental cell line MES-SA, which does not express the mdr1 gene. Poly-SPM had no influence on the IC50 of non-MDR chemotherapeutic agents such as cisplatin. The modulation studies correlated with the ability of Poly-SPM to reverse the cellular accumulation defect of [3H]-VBL and [3H]-TAX in the Dx5 but not MES-SA cells. Pretreatment of the Dx5 cell with alpha-difluoromethylornithine (DFMO at 2 and 5 microM) for 24 h increased the function of the MDR transporter to further decrease the cellular accumulation of VBL and TAX when compared to untreated cells. DFMO pretreatment is known to upregulate the polyamine transporter(s). These findings show that, in addition to inhibiting polyamine transport, Poly-SPM reverses MDR in Dx5 cells, suggesting a potential relationship between the polyamine influx transporter and the MDR efflux pump. This potential functional link between the polyamine influx transporter(s) and the MDR efflux transporter (P-glycoprotein) offers a novel approach to inhibiting this form of drug resistance.

Cell-surface oligosaccharides can function as ligands for intercellular adhesion receptors, matrix proteins, and growth factors. We report that human neonatal and adult epidermal keratinocytes (KC) express sialyl Lewis X [s-Le(x); SA alpha 2-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3R], a ligand for endothelial and platelet selectins. Freshly isolated or cultured KC bind FH6 monoclonal antibody (MoAb), which is specific for s-Le(x)-containing oligosaccharides. The relevant epitope is bona fide s-Le(x), because sialidase treatment of KC suspensions abrogates FH6 binding while generating de novo KC reactivity with anti-Le(x). KC stained in ice-cold suspension display a knobby membrane distribution of s-Le(x) detectable by immunofluorescence microscopy. As others have reported, FH6 appeared not to bind KC in perpendicular skin sections. However, basal KC in intact epidermal sheets exhibited obvious "honeycomb" reactivity with FH6 when stained and viewed en face, suggesting that s-Le(x) in intact epidermis may occur in bands that parallel the major tissue axis. FH6 specifically immunoprecipitated proteins of Mr 34 kd, 44 kd, and 56 kd from [35S]-labeled KC, and anti-Le(x) precipitated similar proteins from sialidase-treated KC. The enzymatic basis for KC s-Le(x) expression was studied by analyzing acceptor specificities and other properties of KC fucosyltransferases. Results indicate that KC express both Lewis- and myeloid-type alpha 1-3fucosyltransferases. KC s-Le(x) could be an important element of the epithelial milieu, because both epithelial cells and immune cells that home to epithelia express s-Le(x) and related structures, and because KC s-Le(x) is well positioned for selectin-mediated platelet binding after trans-cutaneous wounding. The apparent distributions of s-Le(x) in epidermis and on isolated KC are compatible with a functional role for s-Le(x) in these intercellular interactions.

We gauged the internal energy transfer for two dissociative ion decomposition channels in matrix-assisted laser desorption ionization (MALDI) using the benzyltriphenylphosphonium (BTP) thermometer ion [PhCH 2PPh 3] (+). Common MALDI matrixes [alpha-cyano-4-hydroxycinnamic acid (CHCA), 3,5-dimethoxy-4-hydroxycinnamic acid (sinapinic acid, SA), and 2,5-dihydroxycinnamic acid (DHB)] were studied with nitrogen laser (4 ns pulse length) and mode-locked 3 x omega Nd:YAG laser (22 ps pulse length) excitation. Despite the higher fluence required to initiate fragmentation, BTP ions indicated lower internal energy transfer with the picosecond laser in all three matrixes. These differences can be rationalized in terms of phase explosion induced by the nanosecond laser vs a stress-confinement-driven desorption mechanism for the picosecond laser. For the two ion production channels of the BTP thermometer ion, breaking a single bond can result in the formation of benzyl/tropylium ions, F1, or triphenylphosphine ions, F2. In SA and DHB, as well as in CHCA at low fluence levels, the efficiency of these channels (expressed by the branching ratio I F1/ I F2) is moderately in favor of producing tropylium ions, 1 < I F1/ I F2 < 6. As the laser fluence is increased, for CHCA, there is a dramatic shift in favor of the tropylium ion production, with I F1/ I F2 approximately 30 for the nanosecond and the picosecond laser, respectively. This change is correlated with the sudden increase in the BTP internal energies in CHCA in the same laser fluence range. The large changes observed in internal energy deposition for CHCA with laser fluence can account for its ability to induce fragmentation in peptides more readily than SA and DHB.

The long-range goal of the present study is the development of a general approach for in vivo dosimetry of reactive metabolites of polycyclic aromatic hydrocarbons (PAHs), to be used as a tool in cancer risk assessment. With benzo[a]pyrene (BaP) chosen as indicator and a model of PAHs this study aims at the development of a method for the determination of adducts to histidine (His) in hemoglobin (Hb) and serum albumin (SA) of reactive metabolites of BaP. The predominantly mutagenic metabolite of BaP has been shown to be a diolepoxide isomer, +(anti)r-7, t-8-dihydroxy-t-9,10-epoxy-7,8, 9,10-tetrahydrobenzo[a]pyrene (+BPDE). In comparison with other methods for protein degradation, hydrazinolysis was found to be sufficiently effective and mild. The His adduct isolated after protein hydrazinolysis, with protection by tert-butyloxycarbonyl (Boc) of the hydrazide and alpha-amino groups, was shown to be N(im)- +/- (r-7, t-8, t-9-trihydroxy-7, 8, 9, 10-tetrahydrobenzo[a]pyren-c-10-yl)-N(alpha), N(2)-bis(tert-butyloxycarbonyl)-L-histidinehydrazide. Isomers of this compound, used as references, were synthesized and characterized by liquid chromatography/tandem mass spectrometry (LC/MS/MS). Adducts in Hb and SA from in vitro treatment with BPDE were characterized after hydrazinolysis by HPLC-UV/MS, muHPLC/MS/MS and gas chromatography/mass spectrometry (GC/MS). Approximately 70 and 10% of the isolated BPDE adducts from SA and Hb, respectively, were His adducts. Other products were released as BaP tetrols and BaP triols. For the purpose of enrichment/purification of BPDE-His adducts, C(18) and cation exchange solid phase extraction (SPE) were utilized. The sensitivity obtained by this new approach, based on hydrazinolysis of protein, enrichment by SPE and analysis with muHPLC/MS/MS (APCI), is in the low-fmole range.

Microtubules play an essential role in cell division. Little is known about possible variations of total tubulin and tubulin isotype expression during the cell cycle. We analyzed the total tubulin content, tubulin polymerization status and tubulin isotype content in resting and dividing human K562 leukemic cells and human MES-SA sarcoma cells. Although the total cellular tubulin content increases as the cells progress toward mitosis, the total tubulin/total protein ratio is stable during the cell cycle. Reverse transcriptase-polymerase chain reaction was applied to analyze the levels of expression of alpha, beta, and gamma-tubulin isotypes. Whereas alpha-tubulin isotype and gamma-tubulin transcripts were found to be expressed at constant levels throughout the cell cycle, some of the beta-tubulin isotype transcripts were found to be more highly expressed in dividing then in resting cells. Both of the class IV beta-tubulin isotype transcripts (human 5 beta and beta 2, Class IVa and IVb, respectively) were expressed in dividing K562 and MES-SA cells at twice the levels found in resting cells. Increased expression of the class IV isotype proteins in dividing cells was confirmed by immunoblotting, both in K562 and in MES-SA cells. A larger fraction of total cell tubulin was found to be polymerized in dividing cells (36-40%) than in resting cells (27-30%). The degree of polymerization of class IV tubulin in dividing and resting cells was similar to that of total tubulin. These results show that total tubulin is expressed as constant levels throughout the cell cycle but that the degree of polymerization is increased as cells are committed to division. The relative overexpression of the two class IV beta-tubulin isotypes in dividing cells suggests functional specificity for these isotypes and a regulatory role of these isotypes on the microtubule network during mitosis.

Synovial fluid (SF) mononuclear cells (MNC) from 13 patients with rheumatoid arthritis (RA) and 12 patients with other arthritic diseases (OD) including osteoarthritis (OA), gout and spondyloarthritis (SA) were cultured in the presence of collagen types I and II or lipopolysaccharide (LPS) for 24 h. Interleukin-1 (IL-1), IL-6 and tumor necrosis factor-alpha (TNF-alpha) in the SF and culture supernatants were assayed using ELISA. The results showed that one-half of the RA patients with high SF monocyte count had high SF IL-6 levels that coincided with the high spontaneous release of IL-6 by SF MNC. In the other RA patients with lower SF monocyte count, type II collagen induced significantly higher IL-1 beta than the medium control levels by SF MNC (P < 0.01) or that of the other diseases (P < 0.01). Similarly, type II collagen-induced IL-6 and TNF-alpha production rose significantly (P < 0.01) from SF MNC of RA but less from OD (P < 0.05). In addition, type I collagen could also induce IL-1, IL-6 and TNF-alpha in these samples from RA and OD patients but was less potent than type II collagen. Our results indicate that collagen-induced cytokines may be important in the pathogenesis of the disease.

The aim of this study was to observe whether acetylsalicylic acid (ASA) had different effects in both sexes. Out of the ischemic stroke patients who were admitted to the National Taiwan University Hospital (NTUH), those who had not taken ASA or ASA-like drugs for more than 2 weeks were selected for this study. For the diagnosis of ischemic stroke, computed tomography (CT) of the brain was performed in all cases, and for differential diagnosis, other necessary procedures were employed in a few cases. The serum salicylate (SA) level was measured by Trinder's method, thromboxane B2 (TXB2) and 6-keto-PGF1 alpha by radioimmunoassay, threshold concentration of adenosine diphosphate (ADP) by Born's method, and circulating platelet aggregates (CPA) by Wu and Hoak's method. The present study showed that the means of serum SA levels after administration of the same dose of ASA were not significantly different between the two sexes. After ingestion of ASA, a single dose of 75 mg, 300 mg or 600 mg, or 300 mg 4 times a day, mean plasma TXB2 levels were significantly suppressed and mean threshold concentrations of ADP were significantly elevated in the two sexes. After administration of above-mentioned various doses of ASA, the abnormally high plasma TXB2 levels and abnormally low threshold concentrations of ADP and CPA ratios were significantly normalized in both male and female patients. Plasma 6-keto-PGF1 alpha levels were not influenced by ingestion of ASA 75 mg, but significantly depressed by administration of ASA 300 mg in both sexes. There were no sex differences in the antiplatelet effect of ASA in this experiment.

This ex vivo study was performed to evaluate the anti-platelet and anti-thrombogenic potential of shikimic acid (SA), a plant phenolic metabolite. Fasting blood samples were collected from 22 sedentary participants to analyse the effect of varying concentrations of SA (0.1 mM, 0.2 mM, 0.5 mM, 1 mM and 2 mM) on platelet surface-marker expression, platelet aggregation and biomarkers of thrombogenesis. Monocyte-platelet aggregates (CD14/CD42b) and platelet endothelial cell adhesion molecule-1 (PECAM-1 or CD31), effective indicators of thrombus formation were evaluated. Procaspase-activating compound 1 (PAC-1) and P-selectin or CD62P were used to assess platelet activation-related thrombogenesis. Adenosine diphosphate (ADP) was used to stimulate the P2Y1/P2Y12 pathway of platelet activation to mimic the in vivo thrombogenic pathway. Platelet aggregation studies utilised both ADP and collagen as exogenous platelet agonists to target both P2Y1/P2Y12 and GPVI pathways of thrombus formation. It was observed with flow cytometry that SA produced a significant antiplatelet effect on PAC-1 (p = 0.03 at 2 mM) and CD62P (p = 0.017, p = 0.036 at 1 mM and 2 mM respectively) expression in addition to lowering monocyte-platelet aggregate formation (p = 0.013, p < 0.01 and p < 0.01 at 0.5 mM, 1 mM and 2 mM respectively). SA at 1 mM concentration reduced PECAM-1 expression (p = 0.035), signifying a reduction to endothelial leucocyte migration during thrombus growth. SA did not demonstrate a platelet aggregation inhibitory effect by targeting the GPVI collagen pathway but reduced ADP induced platelet aggregation at 2 mM concentration (p < 0.01 at 2 mM). The results suggest that SA, an active metabolite of polyphenol-rich food intake, could play an important role in reducing platelet activation, aggregation related thrombus formation and biomarkers of thrombogenesis in sedentary individuals.

OBJECTIVE

To further validate the Stroke-Adapted Sickness Impact Profile-30 (SA-SIP30) and to determine its responsiveness in a stroke rehabilitation population.

METHODS

Data of 122 communicative stroke patients (mean age 57 years), selected for an inpatient rehabilitation programme, were available. All had suffered different types of stroke. Six months and one year post-stroke, the patients completed the SIP68 plus nine stroke-specific questions from the SIP136, enabling us to derive the SA-SIP30 from the questionnaire. We determined internal consistency, construct and clinical validity and responsiveness of the SA-SIP30. Total, physical and psychosocial dimension scores were calculated.

RESULTS

Internal consistency was moderate to good (alpha>0.68) and correlation between the SIP68 and the SA-SIP30 was high (r>0.85), indicating good construct validity for total score and both dimension scores. Clinical validity assessment showed that total and psychosocial dimensions scores were significantly higher for patients with a cortical infarction compared to respectively subarachnoid haemorrhage and subcortical infarction (p<0.05). Effect sizes for the SA-SIP30 were moderate (between 0.56 and 0.65).

CONCLUSIONS

The SA-SIP30 proved valid and responsive in our stroke rehabilitation population. The major advantages of the SA-SIP30 are the lesser number of items and, therefore, the shorter completion time and the fact that it is a stroke-specific scale to determine health-related functional status.

Localised bone loss in the form of bone erosions and peri-articular osteopenia constitutes an important criteria for the diagnosis of rheumatoid arthritis. In the present study, the effect of Semecarpus anacardium Linn. nut milk extract (SA) on the metabolism of bone turn over has been studied by analyzing various markers of bone turnover and by histological and radiological analysis of the joints in adjuvant arthritis in rats. Arthritis was induced in rats by injecting Freund's complete adjuvant containing 10mg of heat killed mycobacterium tuberculosis in 1 ml paraffin oil (0.1 ml) into the left hind paw of the rat intradermally. After 14 days of induction, SA (150 mg/kg body weight/day) was administered orally by gastric intubations for 14 days. SA significantly reverted the alterations in the bone turnover observed in arthritic animals by modulating the levels of calcium, phosphorus and the activities of the enzymes names tartrate resistant acid phosphatase, acid phosphatase and alkaline phosphatase. The drug increased the bone weights that were found to be decreased during arthritis. Protective effect of SA was also observed by the decrease in the levels and expression of tumour necrosis factor alpha (TNF-alpha) as well as the histopathological and radiological observations. From all these observations it can be concluded that SA possesses strong anti-arthritic property by regulating bone turnover.

Epitestosterone (EpiT) is the 17 alpha-hydroxy epimer of testosterone (T) and a natural steroid metabolite. It has previously been shown to be a 5 alpha-reductase inhibitor. We have studied EpiT as an antiandrogen using the hamster flank organ model. One-centimeter silastic capsules of crystalline T or dihydrotestosterone (DHT) were implanted subcutaneously in female Golden Syrian hamsters to provide continuous androgenic stimulation. After 3 weeks, the pigmented spot was measured and the flank organs were fixed for histologic sectioning. The maximum surface area (SA) from a central section of the sebaceous gland and the diameter of hair follicles were measured using a computerized digitizing tablet. Following T and DHT, respectively, there was a significant increase in pigmented spot size (656/382%), sebaceous gland SA (210/315%), and mean hair follicle diameter (80/56%). A 1-cm capsule of EpiT alone had no androgenic effect. Five- and ten-fold doses of EpiT were implanted with T or DHT. Epitestosterone significantly inhibited the T-dependent stimulation of pigment, sebaceous gland, and hair follicle at either 5- and/or 10-fold excess doses. Additionally, a 10-fold dose of EpiT also inhibited DHT-dependent stimulation of all 3 cutaneous structures. We conclude that EpiT was effective as an antiandrogen and had no intrinsic androgenic activity in the hamster flank organ. It probably functions both as a competitive inhibitor of the androgen receptor and as a 5 alpha-reductase inhibitor. Pigment and sebaceous gland growth were more sensitive than the hair follicle to androgen inhibition by EpiT at the time and doses tested.

The effect of indoramin, a selective alpha 1-adrenoceptor antagonist, on ECG time intervals in normal man was studied. Significant prolongation of QTc was observed after indoramin 100 mg, with no change in QRS duration. In addition, the slope of the RR/QT relationship was changed by indoramin 100 mg. These results suggest that indoramin may have Class III antiarrhythmic activity. If such activity were to affect the SA node, it may explain at least in part the observed lack of reflex tachycardia after indoramin administration.

Non-invasive "hot spot imaging" and localization of necrotic tissue may be helpful for definitive diagnosis of myocardial viability, which is essential for clinical management of ischemic heart disease. We labeled Sennidin A (SA), a naturally occurring median dianthrone compound, with (131)I and evaluated (131)I SA as a potential necrosis-avid diagnostic tracer agent in rat model of reperfused myocardial infarction. Magnetic resonance imaging (MRI) was performed to determine the location and dimension of infarction. (131)I-SA was evaluated in rat model of 24-hour old reperfused myocardial infarction using single-photon emission computed tomography/computed tomography (SPECT/CT), biodistribution, triphenyltetrazolium chloride (TTC) histochemical staining, serial sectional autoradiography and microscopy. Gamma counting revealed high uptake and prolonged retention of (131)I SA in necrotic myocardium and fast clearance from non-targeted tissues. On SPECT/CT images, myocardial infarction was persistently visualized as well-defined hotspots over 24h, which was confirmed by perfect matches of images from post-mortem TTC staining and autoradiography. Radioactivity concentration in infarcted myocardium was over 9 times higher than that of the normal myocardium at 24h. With favorable hydrophilicity and stability, radioiodinated SA may serve as a necrosis-avid diagnostic agent for assessment of myocardial viability.

OBJECTIVE

This paper describes the development of a reliable scale of standards for use in evaluating the progress of the transition from milk to solid food in infants and preschool children. The maturation of chewing and swallowing behavior in infants and young children, which enables processing of solid food, varies, and a scale would assist not only in the instruction of mothers and nurses but also in preventing delay in the introduction of solid food.

METHODS

A range of 159 reference foods were selected on the basis of intake during the period of transition from liquid to solid food. These foods were listed in our previous study, Validity and reliability were tested to create a scale.

METHODS

Foods were selected on the basis of 50% of the subjects studied being able to eat them, and on the food groups classified by cluster analyses using the Varclus procedure of SAS. Validity, of the scale was tested by using Pearson's correlation coefficient between the scale score of selected food items and the total score of all 159 food items. The total score of 159 food items was calculated using the general linear models (GLM) procedure of SAS. Reliability was tested using Cronbach's coefficient alpha.

METHODS

Public health centers in Aomori, Tokyo, Saitama, Nagano, and Okinawa (Japan).

METHODS

Five hundred and eighty healthy mothers and children from 2 to 46 months were randomly selected and 470 (81.0%) completed the study. To avoid regional bias, subjects were drawn from northern to southern prefectures in Japan, namely Aomori, Tokyo, Saitama, Nagano, and Okinawa.

RESULTS

Twenty food items were selected. By analyzing the score correlation using Pearson's correlation coefficient (R =0.97, P <0.001) and GLM (R2 =0.95, P <0.001), it was confirmed that these 20 food items adequately represented the original 159. The reliability was also found to be sufficient (Cronbach's coefficient alpha=0.96).

CONCLUSIONS

The findings demonstrate that a scale of standards for measuring progress in chewing ability can be created using 20 food items. Such a standard will provide a useful basis against which to assess delay of solid food introduction in childhood.

BACKGROUND

Grant-in-Aid for Scientific Research, provided by the Japanese Ministry of Education, Science and Culture, Project No. 07838030.

BACKGROUND

Given the high prevalence of mental health (MH) and substance abuse problems in low-to-middle income countries, the scarcity of MH professionals and the negative impact of psychiatric disorders on caregivers of young children, there is significant need for brief evidence-based screening tools for lay counselors to assist with MH assessment. This study aimed to validate a brief screening tool to assess psychiatric and substance use disorders, the Client Diagnostic Questionnaire (CDQ), in South Africa (SA).

METHODS

Data are from a longitudinal study of health and psychosocial needs in preschool children in SA. Participants included 322 Zulu-speaking, female caregivers. Following procedures of the US CDQ validation study, lay counselors interviewed participants using the translated Zulu CDQ. Subsequently a psychologist conducted a full psychiatric assessment guided by the CDQ questions. Analyses examined sensitivity, specificity and overall accuracy, comparing lay counselor and psychologist assessment.

RESULTS

Sensitivity (73%), specificity (81%) and overall accuracy (79%) were good for the variable indicating presence of 'any diagnosis.' Among those cases identified by the psychologist as having any psychiatric diagnosis, over 70% were correctly identified by lay counselors using the CDQ (i.e., positive predictive value was greater than 70%). The false positive rate was relatively low (19%). Specificity for 'any disorder' (including substance use) and 'any psychiatric disorder' were 81% and 79%.

CONCLUSIONS

The isiZulu CDQ is a sensitive and valid MH diagnostic screener that can be used by lay counselors with limited MH training to identify those in need of treatment and target extremely scarce MH professionals. Copyright © 2016 John Wiley & Sons, Ltd.

UNASSIGNED

South Africa (SA), a country heavily impacted by poverty, HIV and the legacy of Apartheid, has a high prevalence of mental health (MH) and substance abuse problems. In SA and other low-and-middle-income-countries (LMIC) there is a dearth of MH professionals. This study examined use and validity of the Client Diagnostic Questionnaire (CDQ), a brief diagnostic MH screening tool designed for use by lay counselors in HIV-affected populations. Comparing lay counsellor diagnoses on the CDQ to clinician assessment, sensitivity, specificity and overall accuracy were good at the level of 'any diagnosis.' The CDQ can be used effectively in SA and other LMIC with limited MH services to enable appropriate and efficient referral of individuals in primary care settings, supporting caregivers and the children in their care.

OBJECTIVE

To adapt the Cumulative Illness Rating Scale for its use in substance abuse patients (CIRS-SA) and to assess the reliability, internal consistency, and validity of the instrument.

METHODS

One-hundred outpatients of both sexes, 62 men and 38 women, with a mean (SD) age of 32.4 (7.9) years (range 19-57), all of them fulfilling the DSM-IV criteria for any substance abuse disorder. Internal consistency was calculated with Cronbach's alpha coefficient. Test-retest and interrater reliability was assessed with the intraclass correlation coefficient and Wilcoxon z. Validity of the scale was assessed with Kendall's tau correlation coefficient.

RESULTS

The final CIRS-SA version had a total of 13 items. Cronbach's alpha coefficient was 0.57. All intraclass correlation coefficients were above 0.7, and some items showed exact coincidence. The stability of the CIRS-SA scale in a 1-month test re-test reassessment was demonstrated. The CIRS-SA score showed a significant correlation with all consultant scores.

CONCLUSIONS

CIRS-SA is a reliable and valid instrument to assess and to determine systematically the physical condition of substance abusers in whom infections, particularly by the HIV, are highly prevalent.

Immunoglobulin (Ig) heavy chains undergo class switch recombination (CSR) to change the heavy chain isotype from IgM to IgG, A or E. The switch regions are several kilobases long, repetitive, and G-rich on the nontemplate strand. They are also relatively depleted of CpG (also called CG) sites for unknown reasons. Here we use synthetic switch regions at the IgH switch alpha (Sα) locus to test the effect of CpG sites and to try to understand why the IgH switch sequences evolved to be relatively depleted of CpG. We find that even just two CpG sites within an 80 bp synthetic switch repeat iterated 15 times (total switch region length of 1200 bp containing 30 CpG sites) are sufficient to dramatically reduce both Ig CSR and transcription through the switch region from the upstream Iα sterile transcript promoter, which is the promoter that directs transcripts through the Sα region. De novo DNA methylation occurs at the four CpG sites in and around the Iα promoter when each 80 bp Iα switch repeat contains the two CpG sites. Thus, a relatively low density of CpG sites within the switch repeats can induce upstream CpG methylation at the IgH alpha locus, and cause a substantial decrease in transcription from the sterile transcript promoter. This effect is likely the reason that switch regions evolved to contain very few CpG sites. We discuss these findings as they relate to DNA methylation and to Ig CSR.

The effect of succinylacetone (SA), a highly specific inhibitor of ALA-dehydratase and heme synthesis, on hemoglobin (Hb) production, transferrin receptor (TfR), and ferritin expression was analyzed in differentiating Friend leukemia cells (FLC). This compound exerted a pronounced inhibitory effect not only on heme and Hb synthesis, but also on all the remaining above-mentioned parameters. In particular, SA induced: (1) a reduction of the level of alpha-globin mRNA; (2) a decreased number of exposed TfR molecules, without modification of their affinity for the ligand; (3) a reduced level of TfR RNA, without significant change of TfR gene transcription rate; and (4) a lower ferritin content. The addition of exogenous hemin to differentiating FLC exerted opposite effects, and particularly induced an increase of both the number of TfRs and ferritin content. These findings suggest that in erythroid cells optimal heme synthesis is required to coordinately sustain globin chains synthesis and TfR/ferritin production; thus, the intracellular heme level may represent a key regulatory factor in the Hb synthesis pathway.

Investment in and operation of flow control infrastructure such as dams, weirs, and regulators can help increase both the health of regulated river ecosystems and the social values derived from them. This requires high-quality and high-resolution spatiotemporal ecohydrological and socioeconomic information. We developed such an information base for integrated environmental flow management in the River Murray in South Australia (SA). A hydrological model was used to identify spatiotemporal inundation dynamics. River ecosystems were classified and mapped as ecohydrological units. Ecological response models were developed to link three aspects of environmental flows (flood duration, timing, and inter-flood period) to the health responses of 16 ecological components at various life stages. Potential infrastructure investments (flow control regulators and irrigation pump relocation) were located by interpreting LiDAR elevation data, digital orthophotography, and wetland mapping information; and infrastructure costs were quantified using engineering cost models. Social values were quantified at a coarse scale as total economic value based on a national survey of willingness-to-pay for four key ecological assets; and at a local scale using mapped ecosystem service values. This information was integrated using a constrained, nonlinear, mixed-integer, compromise programming optimization model and solved using a stochastic Tabu search algorithm. We tested the model uncertainty and sensitivity using 390 Monte Carlo model runs at varying weights of ecological health vs. social values. Integrating ecohydrological and socioeconomic information identified environmental flow management regimes that efficiently achieved both ecological and social objectives. Using an ecologically weighted efficient and socially weighted efficient scenario, we illustrated model outputs including a suite of cost-effective infrastructure investments and an operational plan for new and existing flow control structures including dam releases, weir height manipulation, and regulator operation on a monthly time step. Both the investments and management regimes differed substantially between the two scenarios, suggesting that the choice of weightings on ecological and social objectives is important. This demonstrates the benefit of integrating high-quality and high-resolution spatiotemporal ecohydrological and socioeconomic information for guiding the investment in and operational management of environmental flows.

An integrated solid-phase spectrophotometry-FIA method is proposed for simultaneous determination of the mixture of saccharin (1,2-benzisothiazol-3(2H)-one-1,1-dioxide; E-954) (SA) and aspartame (N-L-alpha-aspartyl-L-phenylalanine-1-methyl ester; E-951) (AS). The procedure is based on on-line preconcentration of AS on a C18 silica gel minicolumn and separation from SA, followed by measurement, at lambda = 210 nm, of the absorbance of SA which is transiently retained on the adsorbent Sephadex G-25 placed in the flow-through cell of a monochannel FIA setup using pH 3.0 orthophosphoric acid-dihydrogen phosphate buffer, 3.75x10(-3) mol L(-1), as carrier. Subsequent desorption of AS with methanol enables its determination at lambda = 205 nm. With a sampling frequency of 10 h(-1), the applicable concentration range, the detection limit, and the relative standard deviation were from 1.0 to 200.0 microg mL(-1), 0.30 microg mL(-1), and 1.0% (80 microg mL(-1), n = 10), respectively, for SA and from 10.0 to 200.0 microg mL(-1), 1.4 microg mL(-1), and 1.6% (100 microg mL(-1), n = 10) for AS. The method was used to determine the amounts of aspartame and saccharin in sweets and drinks. Recovery was always between 99 and 101%. The method enabled satisfactory determination of blends of SA and AS in low-calorie and dietary products and the results were compared with those from an HPLC reference method.

The objectives of the present investigation were to prepare and characterize starch acetate (SA) with high degree of substitution (dS) and to study its prospect as film-forming agent in a controlled-release multiparticulate drug delivery system. As a part of the development process by quality by design, the objectives also included identification of critical formulation and process variables that affect the release of a drug. SA, a relatively new polymer, was characterized because it showed good film-forming properties. SA with dS 2.9 was synthesized from corn starch by paste disruption technique. It was compared with the raw material, starch, by Fourier transform infrared spectroscopy, X-ray diffraction, and molecular mass analysis. Viscosity of SA solution increased logarithmically with the polymer concentration. At higher polymer concentrations (1.5-5.0%), the solutions showed pseudoplastic behavior. Among the plasticizers tested, triacetin and triethyl citrate yielded free films with acceptable mechanical properties. The glass transition temperature (Tg) of the films could be well controlled by these plasticizers. Unplasticized film showed a Tg of 31.8 degrees C. A trend was found that increase in triacetin concentration in SA films resulted in increase in permeability coefficient for tritiated water. Scanning electron microscopic photographs showed a clear and smooth plasticized film compared to rough unplasticized film. Dyphylline-loaded beads were coated with highly substituted SA to evaluate the main effects of the formulation and process variables on the release of the drug and to figure out the reliability of the screening design. A seven-factor, twelve-run Plackett-Burman screening design was used. The response variables were cumulative percent of drug released in 0.5, 1, 4, 8, and 12 hr. Quantitative evaluation of the design revealed that coating weight gain, plasticizer concentration, and post-drying temperature had greater influence on the drug release than the others. The main effects on drug release after 12 hr decreased in the following order: coating weight gain (-7.81), plasticizer concentration (4.96), postdrying temperature (-2.51), SA concentration (-0.80), inlet temperature (0.51), postdrying time (-0.31), and atomizing pressure (-0.28).

This article describes the functions of a SAS macro and an SPSS syntax that produce common statistics for conventional item analysis including Cronbach's alpha, item difficulty index (p-value or item mean), and item discrimination indices (D-index, point biserial and biserial correlations for dichotomous items and item-total correlation for polytomous items). These programs represent an improvement over the existing SAS and SPSS item analysis routines in terms of completeness and user-friendliness. To promote routine evaluations of item qualities in instrument development of any scale, the programs are available at no charge for interested users. The program codes along with a brief user's manual that contains instructions and examples are downloadable from suen.ed.psu.edu/-pwlei/plei.htm.

The stereotactic aqua stream and aspirator (SAS & A) is a modification of the aqua stream and aspirator (AS & A) designed for stereotactic evacuation of intracerebral hematoma. The needle of the new instrument is inserted into the brain through a burr hole by a conventional stereotactic technique and its tip is directed to the center of the hematoma. The hematoma is broken up with a stream of saline solution from a nozzle. Fragments of clot and fluid can then be aspirated piecemeal through the suction tube. The postoperative CT images show successful evacuation of the hematoma, and the clinical evaluation also showed satisfactory results. This instrument can be used safely, without any complications, early after the stroke.