Investigating differential symptom profiles in major depressive episode with and without generalized anxiety disorder: true co-morbidity or symptom similarity?

BACKGROUND: Large community-based epidemiological surveys have consistently identified high co-morbidity between major depressive episode (MDE) and generalized anxiety disorder (GAD). Some have suggested that this co-morbidity may be artificial and the product of the current diagnostic system. Because of the added direct and indirect costs associated with co-morbidity, it is important to investigate whether methods of diagnostic classification are artificially increasing the level of observed co-morbidity. METHOD: The item response theory (IRT) log-likelihood ratio procedure was used to test for differential item functioning (DIF) of MDE symptoms between respondents with and without a diagnosis of GAD in the 2001-2002 National Epidemiological Survey on Alcohol and Related Conditions (NESARC). RESULTS: The presence of GAD significantly increased the chances of reporting any symptom of MDE, with odds ratios ranging from 2.54 to 5.36. However, there was no indication of significant DIF of MDE symptoms in respondents with and without GAD. CONCLUSIONS: The lack of any significant DIF indicates that cases with GAD do not present with a distinct MDE symptom profile, one that is consistent with the endorsement of symptoms that are conceptually similar in nature between the two disorders, compared to cases without GAD. This does not support the hypothesis that co-morbidity between MDE and GAD is artificially inflated because of the similar symptom criteria required by the current diagnostic system. Instead, MDE and GAD may be thought of as two distinct diagnostic entities that frequently co-occur because of a shared underlying trait.

Ovarian estrogen receptor alpha and beta mRNA expression: impact of development and estrogen.

We tested the hypothesis that ERalpha and ERbeta mRNAs in the rat ovary are regulated during the post-natal period and in immature rats in response to estrogen treatment. Total ovarian ERbeta mRNA was more abundant than ERalpha mRNA and expression of ERbeta increased between post-natal days 4 and 12, coinciding with advancing folliculogenesis and an increase in granulosa cell numbers. In contrast, ERalpha mRNA levels remained relatively constant during this period. In situ hybridisation studies localised both ERalpha and ERbeta to granulosa cells of growing follicles, in 25 day old ovaries, although not all granulosa cells in a follicle or all follicles expressed the ERs. Diethylstilboestrol (DES) administered in vivo to 21 day old rats, for up to 4 days, did not significantly alter the expression of either ER as determined by RT-PCR, despite a 5.5-fold increase in granulosa cell number in these ovaries. In situ hybridisation studies established that DES-treatment down-regulated granulosa cell ER mRNAs. RT-PCR analyses on isolated granulosa cells confirmed that ERalpha was significantly down-regulated by DES. The predominance of ERbeta over ERalpha in the ovary and the regulation of ERbeta mRNA expression during ovarian development, is consistent with an important biological role for ERbeta in granulosa cell proliferation and differentiation.

Production and actions of inhibin and activin during folliculogenesis in the rat.

Evidence to enhance the premise that inhibin and activin are local regulators of ovarian folliculogenesis is presented in this review. Granulosa cells (GC) have been identified as the source of inhibin/activin in the ovary on the basis of mRNA and protein localisation and the measurement of the inhibin forms in GC conditioned media. Expression of the subunit mRNAs changed with follicular development, being maximal in the ovaries of 8-day-old rats, where secondary follicles predominate. The expression of beta subunit mRNAs by GC isolated from diethylstilboestrol (DES)-treated immature rats, was reduced in the absence of any change in alpha subunit mRNA expression. Dimeric inhibin-A, -B and free alpha subunit were produced by ovarian cell cultures prepared from 4- to 12-day-old rats. Inhibin-A production by these cultures was responsive to FSH and TGF-beta, with preantral follicles of day 8 ovaries exerting effects so profound that the inhibin A/alpha subunit ratio increased, most likely due to a stimulation of beta(A) subunit production. In contrast, inhibin-B was not stimulated by TGF-beta until day 8 and FSH until day 12. Fractionation of GC conditioned media revealed a prominence of free alpha subunit and inhibin-A, but little inhibin-B, suggesting that inhibin-B production declines with follicular development. Activin receptor types I and II, Smads 1-8 and betaglycan (beta-glycan) mRNAs were present in the rat ovary and showed distinct patterns of expression between postnatal days 4 and 12. Oocytes and GC localised activin receptor, Smad and beta-glycan proteins, with beta-glycan also present in theca cells (TC). These data indicate that activin/TGF-beta signalling machinery and factors which influence these pathways, are present in the postnatal rat ovary. Our hypothesis that inhibin and activin play important and changing autocrine/paracrine roles in the growth and differentiation of follicles, including the oocyte, has been supported by these studies.

Recruitment and development of the follicle; the roles of the transforming growth factor-beta superfamily.

Peripheral endocrine hormones and local paracrine and autocrine factors contribute, in a coordinated fashion, to the processes of recruitment, development or atresia, selection and ovulation of follicles. Among the local ovarian factors, there is growing evidence from genetic and experimental data that many members of the transforming growth factor (TGFbeta) superfamily have a biological role to play in folliculogenesis. These members include activin, inhibin, TGFbeta, BMP, GDF9 and perhaps MIS. In this review, we discuss the potential roles of the TGFbeta superfamily members, in particular activin, during folliculogenesis. Since the actions of these factors are determined by ligand availability, receptor expression and modulation of their signal transduction pathways, we also collate information on the expression of their signalling components in the follicle. We conclude that the TGFbeta superfamily signalling pathways, in particular activin's pathway, reside in the ovary. Furthermore, follistatin and beta-glycan-components of the accessory binding protein system that modifies activin action-are also present in follicles. In the post-natal rat ovary, the changes in receptor/Smad expression coincide with granulosa cell proliferation and antrum formation. We hypothesise that these pathway components are expressed in a temporal and cell-specific manner to meet the changing demands of cells during follicular development. The analysis of the components of the signal transduction pathways of the TGFbeta family members in populations of defined follicles and the identification of activated pathways in individually stimulated follicles should help clarify the roles of the TGFbeta members in folliculogenesis.

Visceral leishmaniasis in the BALB/c mouse: a comparison of the in vivo activity of five non-ionic surfactant vesicle preparations of sodium stibogluconate.

Five non-ionic surfactants (Surfactants V-IX) were screened for their ability to produce vesicles for the delivery of sodium stibogluconate. Mean vesicle diameter and antimony content were determined prior to in vivo assessment of antiparasitic activity in a mouse model of acute visceral leishmaniasis. V/D suspensions (i.e. stibogluconate loaded vesicles kept in the hydrating drug solution) were more effective against spleen, liver and bone marrow parasites than drug loaded vesicle suspensions that had unentrapped drug removed. A Surfactant IX V/D suspension was the most active antileishmanial preparation causing 74 +/- 10%, 99 +/- 1% and 38 +/- 8% suppression of liver, spleen and bone marrow parasite burdens respectively. Contrary to previous findings, a reduction in splenic and bone marrow parasite burdens was achieved using large vesicles (mean diameter > 800nm). The significance of these results is discussed.

The distribution of free and non-ionic vesicular sodium stibogluconate in the dog.

The pharmacokinetics and tissue distribution of antimony after the administration of sodium stibogluconate in a free form or entrapped in vesicles prepared from non-ionic surfactant were studied in the dog. Animals were given either one or two intravenous bolus injection(s) equivalent to 45 mg Sb kg-1 as free drug or 0.625 or 0.685 mg Sb kg-1 as vesicular drug. Blood samples were taken at various times after dosing and antimony levels in various tissues were determined at 3 h, 48 h and 6 days after dosing. After free stibogluconate antimony clearance from the blood occurred in a rapid elimination phase with a blood half-life of 0.58 +/- 0.08 h. This rapid elimination phase did not occur after vesicular drug. Both drug preparations gave similar antimony levels in the spleen, liver and femur and humerus bone marrow at all time points assessed even though the vesicular dose was one-seventieth of the free drug dose. After the free drug there was marked urinary excretion of antimony and, as a result, increased kidney loading at the expense of other tissue. Vesicle-mediated drug delivery suppressed renal excretion and a much greater proportion of the antimony dose was recovered from tissue than was obtained after free drug. A hypothesis is presented to account for the differences in tissue antimony concentrations produced by the two formulations.

Avoidant personality disorder: empirical support for DSM-IV revisions.

This article attempts to answer three questions about avoidant personality disorder (AVPD): (a) Is it a coherent unidimensional entity, (b) Is the requirement that four or more of the criteria be met in order to make the diagnosis justified, and (c) Are the changes made in DSM-IV supported? Four hundred thirty-four people presenting for treatment for anxiety were assessed with the Personality Disorder Examination. The criteria met were factor analyzed to indicate the unidimensionality of the diagnosis. Measures of internal consistency were calculated to validate the instruction to diagnose AVPD if four or more criteria were observed. Confirmatory factor analysis showed that a single factor was the best fit to the observed pattern of relationships between the seven AVPD criteria. The internal consistency of the seven criteria was moderate (Cronbach's alpha = .76) with a median intercriterion correlation of .29. The data provided good support for the hypothesis that the seven DSM-III-R AVPD criteria assess a single dimension. Three of the criteria did not reflect this factor as highly as the remaining four and these three have either been dropped in DSM-IV or substantially revised.

Genetic control of drug-induced recovery from murine visceral leishmaniasis.

The influence of host genetic background on the response of Leishmania donovani-infected mice to chemotherapy was studied using the H-2d and H-2b haplotypes on a BALB or a B10 genetic background. Animals were treated with free or liposomal sodium stibogluconate and parasite burdens in the liver, spleen and bone marrow were assessed. In all the mouse strains and their congenic derivatives examined, the liver responded best to therapy regardless of drug formulation, whilst the spleen and the bone marrow respectively were increasingly less responsive to chemotherapy. Treatment with free drug was more effective in congenic mice carrying the H-2b haplotype than in those carrying the H-2d haplotype and in mice carrying the same H-2 haplotype, animals from a BALB background were better responders than those from a B10 genetic background. Liposomal drug was more effective than free drug treatment in all four mouse strains and produced a similar significant suppression (> 99%, P < 0.001) in liver parasite burdens to that obtained using a six times greater free drug dose. This liposomal drug dose was more effective than free drug in reducing bone marrow parasite burdens in all four mouse strains and equally (BALB/c mice) or more effective (P < 0.01, BALB/B, B10 and B10.D2 strains) in reducing spleen parasite numbers. Liposomal dependent influences apparent using free sodium stibogluconate. These results are discussed in relation to the genetic factors which are known to control the course of L. donovani infection in mice.

An in-vitro model of intracellular bacterial infection using the murine macrophage cell line J774.2.

A simple model of intracellular bacterial infection based on the ability of the macrophage cell line J774.2 to phagocytose Escherichia coli in a reproducible manner is described. Viable counting of intracellular bacteria and microscopic examination of infected macrophage cultures after treatment with fluorescent E. coli antibody showed that the bacteria multiplied within the J774.2 cells. Viable intracellular bacteria may be used to study the activity of bactericidal and bacteriostatic drugs within the macrophage. The low apparent intracellular bactericidal activity of streptomycin, which was time- and concentration-dependent, accords with a low permeability of the J774.2 macrophages to this antibiotic. An exclusively intracellular infection could be achieved by inactivation of non-phagocytosed extracellular bacteria by streptomycin treatment of infected macrophage cultures. Under appropriate conditions intracellular bacterial viability was unaffected.

Quantification of uptake of liposomal carboxyfluorescein by professional phagocytes in-vitro. A flow microfluorimetric study on the J774 murine macrophage cell line.

Unilamellar egg phosphatidylcholine/cholesterol liposomes containing carboxyfluorescein were prepared by an ether injection method. The ability of cells of the J774.2 murine macrophage cell line to incorporate the liposomal fluorophore during incubation at 37 degrees C was measured by flow microfluorimetry. Liposomes incorporating additional phosphatidylserine or phosphatidic acid were taken up much more avidly than those lacking these phospholipids and the greatest uptake of carboxyfluorescein was observed with the phosphatidylserine species. Calculation of the number of liposomes taken up, greater than or equal to 0.2% of the number given, showed that this was an inefficient process. However these uptake data support previous findings based on the intracellular bactericidal activity of liposomal antibiotics determined in an identical in-vitro system.

The therapeutic effect of sodium stibogluconate in BALB/c mice infected with Leishmania donovani is organ-dependent.

A study of the antileishmanial efficacy of sodium stibogluconate was carried out in BALB/c mice. The drug was administered to Leishmania donovani-infected animals on days 7 and 8 post-infection in one of three forms; free (40-50 mg Sbv Kg-1), liposomal, or niosomal (6.4-8.0 mg Sbv Kg-1) drug. On day 14 post-infection counts of the number of parasites present in the liver, spleen and bone marrow of treated and control animals showed that although all three drug preparations significantly reduced parasite numbers in the liver (approximately equal to 99% suppression) they had little effect on those residing in the spleen or bone marrow. The carrier forms of the drug were therefore significantly more effective than free drug in reducing liver parasite burdens. Increasing the concentration and the number of doses of free drug (maximum of 500 mg Sbv Kg-1), and reducing the size of the vesicles used to deliver the drug had a minimal effect on parasite numbers in the spleen and bone marrow. It is proposed that because of the resistance of spleen and bone marrow parasites to drug therapy, the BALB/c mouse infected with L. donovani provides an excellent model system for the study of drug delivery to these deeper tissue sites.

Comparison of the efficacy of free and non-ionic-surfactant vesicular formulations of paromomycin in a murine model of visceral leishmaniasis.

Non-ionic-surfactant vesicular (NIV) formulations of paromomycin have been tested in-vitro and in-vivo for their activity against Leishmania donovani. Production of NIV was dependent both on the surfactant used and on the concentration of paromomycin; only two of the surfactants studied formed vesicles at the highest paromomycin concentration (9 mg mL(-1)). At surfactant-lipid concentrations > or = 1.5 mM, suspensions of NIV (drug- or glucose-loaded) were cytotoxic to macrophages infected with L. donovani; high levels of nitrite were produced in cell supernatants. At surfactant-lipid concentrations < 1.5 mM, drug-loaded NIV were more effective than the same dose of free drug, in terms of the percentage of cells infected and the number of parasites/cell. At surfactant-lipid concentrations < or = 0.15 mM, drug-loaded NIV were ineffective in-vitro. In-vivo, treatment with decaethylene glycol mono n-hexadecyl ether paromomycin NIV was more effective than hexaethylene glycol mono n-hexadecyl ether paromomycin NIV, in terms of suppression of liver and spleen parasite burdens. Against liver parasites, both types of paromomycin-loaded NIV were more effective than free drug. Neither the NIV nor free forms of paromomycin caused significant suppression of bone-marrow parasites. The study shows that entrapment of paromomycin in NIV can be used to increase its antileishmanial activity in-vitro and in-vivo.

Non-ionic surfactant vesicles, niosomes, as a delivery system for the anti-leishmanial drug, sodium stibogluconate.

Liver and serum concentrations of antimony in the mouse have been determined after administration of sodium stibogluconate in the free, liposomal and niosomal form. High liver and low serum values were attained by the use of both vesicular formulations. Niosomal sodium stibogluconate was shown to be more active than free drug against experimental murine visceral leishmaniasis, an effect apparently dependent on maintaining high drug levels in the infected reticuloendothelial system.

Vesicular systems (niosomes and liposomes) for delivery of sodium stibogluconate in experimental murine visceral leishmaniasis.

Suppression of Leishmania donovani liver amastigotes by sodium stibogluconate has been determined in a murine model of experimental visceral leishmaniasis. Niosomal and liposomal drug formulations were equiactive and both increased drug efficacy by an order of magnitude compared with that of free drug. Niosomes containing 30 mol % cholesterol were prepared from three different non-ionic surfactants and no significant difference in activity was detected among the different drug-loaded niosomes. Both negatively charged and neutral vesicles were found to be equally effective. However, vesicle cholesterol content had a slight influence on the antiparasitic activity of the drug-loaded niosomes. Empty vesicles produced a dose-dependent parasite suppression for all vesicles studied. Studies of antimony distribution in the mouse using neutron activation analysis showed high liver levels after i.v. administration of the carrier forms of the drug.

Visceral leishmaniasis: drug carrier system characteristics and the ability to clear parasites from the liver, spleen and bone marrow in Leishmania donovani infected BALB/c mice.

The efficacy of various sodium stibogluconate formulations against Leishmania donovani has been investigated using a BALB/c mouse model of visceral leishmaniasis. Only one therapy, multiple dosing with drug loaded sonicated vesicles, liposomes or niosomes, was found to be effective against parasites in the liver, spleen and bone marrow. Other treatments significantly reduced parasite liver burdens but either failed to effect spleen and bone marrow parasites, or were effective but toxic. Prophylactic treatment with sodium stibogluconate preparations, six days before infection, reduced parasite multiplication in the liver (free, niosomal and liposomal drug) and the spleen (sonicated, drug loaded niosomes only), but had no suppressive effect on bone marrow parasite burdens compared with controls. These results indicate that in-vivo sodium stibogluconate persists in some compartments at parasiticidal concentrations and that failure to reach this concentration at some sites of infection such as bone marrow, is the cause of treatment failure and relapse.

The preparation and properties of niosomes--non-ionic surfactant vesicles.

Vesicles were prepared on hydration of a mixture of a single or double alkyl-chain, non-ionic surfactant with cholesterol. These vesicles, or 'niosomes', are capable of entrapping and retaining water soluble solutes such as carboxyfluorescein, are osmotically active and can be formulated to release entrapped solute slowly. The physical characteristics of the vesicles were found to be dependent on the method of production and three such methods, based on liposome technology, are described. The vesicles have been characterized by photon correlation spectroscopy, freeze fracture electron micrography, measurement of solute entrapment efficiency, and solute release rates. Vesicular forms of the single chain surfactant which could be formed under certain conditions in the absence of cholesterol are also described.

Biodegradable microspheres: polyacryl starch microparticles as a delivery system for the antileishmanial drug, sodium stibogluconate.

Liver parasite burdens of Leishmania donovani in the mouse have been determined after treatment with intravenous administration of sodium stibogluconate in the free or carrier form. The carrier form, in which the drug was covalently bound to polyacryl starch microparticles, was up to 100x more effective than the free form in this murine model of visceral leishmaniasis. Empty microparticles had no effect on liver parasite burdens and the enhanced in-vivo antileishmanial activity of the carrier form of the drug was apparently due to passive drug delivery to the infected liver.

An Australian survey of the procedures used for the treatment of opiate users.

Drug and alcohol agencies across Australia were asked to describe the services they offer to opiate users. Of the 284 agencies identified as providing treatment, 229 (81%) responded. A standard assessment procedure was used in 72% of agencies. Eighty (35%) agencies offered detoxification and had assisted 7883 clients with detoxification in the 12 months prior to March 1990. Methadone maintenance was offered in 20% of agencies with 5234 clients currently receiving this treatment. Daily doses of methadone in the range of 40-80 mg were described for most (51%) clients receiving methadone and concurrent counselling was provided in 45% of cases. Standard psychosocial interventions were provided by 60% of agencies. Out-patient or non-residential settings were most common (36%), with residential therapeutic communities being the setting for 21% of programmes. Supportive counselling was the most commonly used individual approach, and cognitive-behavioural or 12-step approaches were the most commonly used group approaches. Brief support or referral to Nar-Anon were the most popular family interventions. Procedures aimed at reducing the risk of HIV were in place at 85% of agencies. These findings are discussed in light of research evidence. Briefly, there is a diversity of treatment options available from different treatment agencies which is not reflected within the agencies, little aftercare is offered despite high rates of relapse, and doses of methadone are lower than has been found to be optimal.

Characteristics of clients receiving treatment in Australian drug and alcohol agencies: a national census.

The first Australian national census of clients of drug and alcohol treatment agencies was undertaken to document the characteristics of clients attending these facilities. Of the agencies surveyed nationwide, 431 (85.2%) completed and returned census forms describing demographic and drug use details of their clients. A total of 6175 clients were reported to have received services (beyond methadone maintenance alone) in the responding agencies on the census day. Ninety percent of the clients seen were substance users, and 10% of clients were relatives or friends of a substance user. The mean age of the clients was 34 years and two-thirds were male. The majority were Australian born, with 10% described as Aborigines or Torres Strait Islanders. The majority of the clients were not in paid employment. Fifty-five percent of all clients received treatments services on a non-residential basis. The most frequent presenting drug problem reported was alcohol, followed by opiates and tobacco. Thirty-three percent of the substance users were reported to have injected illicit drugs in the past 12 months.

Meta-analytic review of the efficacy of smoking cessation interventions.

A meta-analysis of randomized and controlled evaluations of the efficacy of smoking cessation interventions compared 146 estimates of the difference in abstinence rates between treated and control conditions (effect sizes) from 85 publications. Simple advice to quit and other brief intervention techniques, nicotine chewing gum and behavioural techniques were all found to be significantly better than relevant control conditions in promoting abstinence, although the results were not homogeneous. In five studies of acupuncture compared with control, consistent results were found showing no benefit for acupuncture.