Belimumab restores Treg/Th17 balance in patients with refractory systemic lupus erythematosus.

Belimumab, a specific inhibitor of the soluble B lymphocyte stimulator (BlyS), is the first biological drug approved by the United States Food and Drug Administration for the treatment of patients with active systemic lupus erythematosus (SLE) refractory to standard therapy. Given that an imbalance between regulatory T cells (Treg) and interleukin (IL)-17A-secreting T cells (Th17) has been reported in various autoimmune disorders, we assessed the frequency of both Treg and Th17 peripheral blood populations before and after belimumab administration in 20 patients with active SLE refractory to standard therapy. After six months of treatment, the mean SELENA-SLEDAI score as well as the mean anti-double-stranded DNA antibody titers were significantly decreased. In addition, we observed a significant increase in Treg percentages and a parallel, significant decrease in Th17 percentages, accompanied by significantly reduced serum levels of IL-21. In vitro studies showed that Treg purified from belimumab-treated patients were fully functional and displayed a suppressor function similar to that of Treg purified from healthy donors. Belimumab can restore Treg/Th17 balance in SLE patients with uncontrolled disease activity, and this results in decreased flare rate and reduced glucocorticoid dosage.

Therapeutic targeting of the mTOR-signalling pathway in cancer: benefits and limitations.

The mammalian target of rapamycin (mTOR) plays an important role in the regulation of protein translation, cell growth and metabolism. The mTOR protein forms two distinct multi-subunit complexes: mTORC1 and mTORC2. The mTORC1 complex is activated by diverse stimuli, such as growth factors, nutrients, energy and stress signals; and essential signalling pathways, such as PI3K and MAPK, in order to control cell growth, proliferation and survival. mTORC1 also activates S6K1 and 4EBP1, which are involved in mRNA translation. The mTORC2 complex is resistant to rapamycin inhibitory activity and is generally insensitive to nutrient- and energy-dependent signals. It activates PKC-α and Akt and regulates the actin cytoskeleton. Deregulation of the mTOR-signalling pathway (PI3K amplification/mutation, PTEN loss of function, Akt overexpression, and S6K1, 4EBP1 and eIF4E overexpression) is common in cancer, and alterations in components of the mTOR pathway have a major role in tumour progression. Therefore, mTOR is an appealing therapeutic target in many tumours. Here we summarize the upstream regulators and downstream effectors of the mTORC1 and mTORC2 pathways, the role of mTOR in cancer, and the potential therapeutic values and issues related to the novel agents targeting the mTOR-signalling pathway.

Are retinoids a promise for Alzheimer's disease management?

Retinoids regulate several physiological and pathological processes through the interaction with nuclear receptors. Retinoid-associated signaling which plays an essential role in neurodevelopment appears to remain active in the adult central nervous system (CNS), thus assuming a high significance in the context of neurodegeneration, and indeed retinoid analogs are thought to be promising therapeutic agents for the treatment of neurodegenerative disorders. The ability of retinoids to exert antioxidant effects, inhibit amyloid-ß (Aß) deposits and likely Aß-induced mitochondrial dysfunction, tau hyperphosphorylation, Aß-induced IL6 production and neuroinflammation, increase survival in hippocampal neurons, and reverse cognitive deficits in animal models of Alzheimer's disease (AD) is discussed. Although retinoids with their multi-target activity are revealing to be promising for management of AD which is a multifaceted biochemical phenomenon, timing as well as appropriate dosage and safety remain, however, a challenge. The end-stage lesions, namely senile plaques and neurofibrillary tangles, are, at present, considered an adaptive response to oxidative stress underlying AD, thus paradoxically late administration of retinoids could even suppress a protective mechanism by inhibiting Aß deposits.

Retinoids: novel immunomodulators and tumour-suppressive agents?

Retinoids play important roles in the transcriptional activity of normal, degenerative and tumour cells. Retinoid analogues may be promising therapeutic agents for the treatment of immune disorders as different as type I diabetes and systemic lupus erythematosus. In addition, the use of retinoids in cancer treatment has progressed significantly in the last two decades; thus, numerous retinoid compounds have been synthesized and tested. In this paper, the actual or potential use of retinoids as immunomodulators or tumour-suppressive agents is discussed.

Towards monitoring real-time cellular response using an integrated microfluidics-matrix assisted laser desorption ionisation/nanoelectrospray ionisation-ion mobility-mass spectrometry platform.

The combination of microfluidic cell trapping devices with ion mobility-mass spectrometry offers the potential for elucidating in real time the dynamic responses of small populations of cells to paracrine signals, changes in metabolite levels and delivery of drugs and toxins. Preliminary experiments examining peptides in methanol and recording the interactions of yeast and Jurkat cells with their superfusate have identified instrumental set-up and control parameters and online desalting procedures. Numerous initial experiments demonstrate and validate this new instrumental platform. Future outlooks and potential applications are addressed, specifically how this instrumentation may be used for fully automated systems biology studies of the significantly interdependent, dynamic internal workings of cellular metabolic and signalling pathways.

The impact of professional and social network support on the burden of families of patients with schizophrenia in Italy.

OBJECTIVE: (a) To explore burden related to caregiving and support received from professionals and social network in relatives of patients with schizophrenia in Northern, Central and Southern Italy; and (b) to test whether a higher level of family burden is associated with a lower level of professional and social network support. METHOD: Seven hundred and nine patients with schizophrenia and their key-relatives were consecutively recruited in 30 Italian mental health departments. Data were collected on: (a) patients' clinical status and levels of disability; (b) relatives' burden, social and professional support; (c) interventions received by patients and their families. RESULTS: Family burden was found lower in Northern Italy. However, after controlling for psychosocial interventions, differences in family burden among the three geographical areas disappeared. Family burden was associated with patients' levels of disability and manic/hostility symptoms, and with professional and social network support received by the family. CONCLUSION: Professional and social network support represent crucial resources to reduce family burden in schizophrenia.

A multicenter Italian study of patients' relatives' beliefs about schizophrenia.

This study explored patients' relatives' beliefs about schizophrenia in Italy, a country with a long history of community-based psychiatry. The study was conducted among 709 key relatives of patients with schizophrenia who were recruited in 30 mental health departments. Relatives seemed to believe that mental hospitals are not appropriate places in which to treat their loved ones, even though most of them felt that they were alone in coping with the burden of their ill relative's disease. Most of the relatives agreed that people who have schizophrenia should be allowed to vote, but they appeared to have a stricter attitude toward affective rights, such as the patient's right to get married and have children, and tended to perceive their loved ones as being subject to social discrimination. The findings of this study may be useful for developing psychoeducational intervention and sensitization campaigns for the general population.

[The condition of families of patients with schizophrenia in Italy: burden, social network and professional support]. / La situazione delle famiglie dei pazienti con schizofrenia in Italia: carico familiare, risposte dei SSM, sostegno sociale.

OBJECTIVE: This study aimed to describe: a) the interventions received by patients with schizophrenia attending Italian mental health services (MHS); b) the relatives' burden and social network and the professional support received by the families. DESIGN: The study has been carried out in 30 MHS, randomly selected and stratified by geographic areas and population density. 25 patients with a DSM-IV diagnosis of schizophrenia and 25 relatives were recruited in each MHS. Family burden was evaluated in relation to: a) geographic area; b) interventions received by the patients; c) social and professional support received by the families. MAIN OUTCOME MEASURES: a) patients: Brief Psychiatric Rating Scale (BPRS) and Disability Assessment Interview (AD); b) key-relatives: Family Problems Questionnaire (QPF) and Social Network Questionnaire (QRS); c) interventions received by the patients and their families: Scheda di Rilevazione degli Interventi--Pattern of Care Schedule (SRI). RESULTS: Data on 709 patients and their key-relatives were collected. In the two months preceding the data collection, 35% of patients attended rehabilitative programmes; 80% of the families were in regular contact with the MHS and 8% received family psychoeducational interventions. Family burden was higher in Southern than in Central and Northern Italy. This difference disappeared when rehabilitative interventions and family support were provided. CONCLUSIONS: This study highlights that the situation of the families of patients with schizophrenia is more burdensome in Southern Italy and is greatly influenced by the type of interventions provided by the MHS.

[Causes and psychosocial consequences of schizophrenia: the opinions of patient's relatives]. / Cause e conseguenze psicosociali della schizofrenia: le opinioni dei familiari dei pazienti. Gruppo di Lavoro.

OBJECTIVE: Description of the opinions on schizophrenia and its psychosocial consequences in a sample of relatives of patients with schizophrenia, recruited in 30 Italian Mental Health Centres (MHC), stratified by geographic areas and population density. DESIGN: Cross-sectional study on key-relatives of clinically stable patients with a DSM-IV diagnosis of schizophrenia. Evaluation of relatives' opinions on schizophrenia and its psychosocial consequences, in relation to: a) patient's clinical and family's socio-demographic variables; b) geographic areas and population density. SETTING: 30 Italian MHC randomised and stratified by geographic areas (Northern, Central, Southern Italy) and population density (> 100,000 inhabitants; between 100,000 and 25,000 inhabitants; < 25,000 inhabitants). Main outcome measures--Patient's clinical status and social functioning: Brief Psychiatric Rating Scale (BPRS) and Disability Assessment interview (AD). Interventions provided to patient and his/her family: Scheda di Rilevazione degli Interventi--Pattern of Care Schedule (SRI). Relatives' opinions on schizophrenia and its psychosocial consequences: Questionnaire on the Opinions of the Family (QOF). RESULTS: Data on 709 patients and their key-relatives were collected. Relatives' pessimistic opinions on social competence of patients with schizophrenia were found to be associated with: high levels of patient's disability, hostility and negative symptoms, relatives' knowledge of patient's diagnosis of schizophrenia, residence in a medium or low population density area, low levels of relative's education. Relatives' pessimistic opinions on social restrictions imposed by schizophrenia were found to be associated with high levels of disability and high number of hospital admissions in the patient and older age in the relative. CONCLUSIONS: The results of this study emphasise the need to: a) provide families with educational interventions covering not only clinical aspects of schizophrenia, but also those concerning disability and psychosocial consequences of the disorder; b) plan educational campaigns on mental illnesses which take into account the socio-cultural characteristics of the target populations.

Family burden and coping strategies in schizophrenia: 1-year follow-up data from the BIOMED I study.

BACKGROUND: To date, only few data are available on how family burden in schizophrenia changes over time. In addition, no study has explored how such factors as coping styles and social support influence burden over time. This paper presents the 1-year follow-up data from the BIOMED I study on family burden and coping strategies in schizophrenia. METHODS: A sample of 159 relatives of patients with schizophrenia living in five European countries was followed up prospectively for 1 year with regard to burden and coping strategies, using validated questionnaires. RESULTS: In the sample as a whole, the burden was stable. A reduction of family burden over time was found among relatives who adopted less emotion-focused coping strategies and received more practical support from their social network. In addition, family burden decreased in relation to the improvement of patient's social functioning. CONCLUSIONS: When relatives of patients with schizophrenia are able to improve their coping strategies, it is possible for burden to be reduced even after several years. This points to the necessity to provide families of chronic psychotic patients with psychoeducational interventions emphasising the adoption of an effective coping style.

A new questionnaire assessing the opinions of the relatives of patients with schizophrenia on the causes and social consequences of the disorder: reliability and validity.

This paper describes the development and the validation of a questionnaire assessing the opinions of the relatives of patients with schizophrenia on the causes and the social consequences of the disorder. The final version of the questionnaire includes 28 items, grouped in four sub-scales (social restrictions, social distance, utility of treatments, biopsychosocial causes of schizophrenia), whose intra-rater reliability ranges from 0.36 to 0.84. Cronbach's alpha coefficient, which tests the content validity of the sub-scales, ranges from 0.56 to 0.66. Factor analysis identifies two factors (opinions on social consequences and the utility of treatments, and the beliefs on the causes of schizophrenia), which account for the 73% of the total variance. This questionnaire may be particularly useful to assess relatives'beliefs about schizophrenia and to target psychosocial interventions in the families of patients with schizophrenia.

Mechanism of inhibition of DNA (cytosine C5)-methyltransferases by oligodeoxyribonucleotides containing 5,6-dihydro-5-azacytosine.

A key step in the predicted mechanism of enzymatic transfer of methyl groups from S-adenosyl-l-methionine (AdoMet) to cytosine residues in DNA is the transient formation of a dihydrocytosine intermediate covalently linked to cysteine in the active site of a DNA (cytosine C5)-methyltransferase (DNA C5-MTase). Crystallographic analysis of complexes formed by HhaI methyltransferase (M.HhaI), AdoMet and a target oligodeoxyribonucleotide containing 5-fluorocytosine confirmed the existence of this dihydrocytosine intermediate. Based on the premise that 5,6-dihydro-5-azacytosine (DZCyt), a cytosine analog with an sp3-hybridized carbon (CH2) at position 6 and an NH group at position 5, could mimic the non-aromatic character of the cytosine ring in this transition state, we synthesized a series of synthetic substrates for DNA C5-MTase containing DZCyt. Substitution of DZCyt for target cytosines in C-G dinucleotides of single-stranded or double-stranded oligodeoxyribonucleotide substrates led to complete inhibition of methylation by murine DNA C5-MTase. Substitution of DZCyt for the target cytosine in G-C-G-C sites in double-stranded oligodeoxyribonucleotides had a similar effect on methylation by M. HhaI. Oligodeoxyribonucleotides containing DZCyt formed a tight but reversible complex with M.HhaI, and were consistently more potent as inhibitors of DNA methylation than oligodeoxyribonucleotides identical in sequence containing 5-fluorocytosine. Crystallographic analysis of a ternary complex involving M.HhaI, S-adenosyl-l-homocysteine and a double-stranded 13-mer oligodeoxyribonucleotide containing DZCyt at the target position showed that the analog is flipped out of the DNA helix in the same manner as cytosine, 5-methylcytosine, and 5-fluorocytosine. However, no formation of a covalent bond was detected between the sulfur atom of the catalytic site nucleophile, cysteine 81, and the pyrimidine C6 carbon. These results indicate that DZCyt can occupy the active site of M.HhaI as a transition state mimic and, because of the high degree of affinity of its interaction with the enzyme, it can act as a potent inhibitor of methylation.

Burden on the families of patients with schizophrenia: results of the BIOMED I study.

The burden, the coping strategies and the social network of a sample of 236 relatives of patients with schizophrenia, living in five European countries, were explored by well-validated assessment instruments. In all centres, relatives experienced higher levels of burden when they had poor coping resources and reduced social support. Relatives in Mediterranean centres, who reported lower levels of social support, were more resigned, and more often used spiritual help as a coping strategy. These data indicate that family burden and coping strategies can be influenced by cultural factors and suggest that family interventions should have also a social focus, aiming to increase the family social network and to reduce stigma.

Social and clinical factors influencing the choice of coping strategies in relatives of patients with schizophrenia: results of the BIOMED I study.

The impact of social and clinical factors on the choice of coping strategies of a sample of 236 relatives of patients with schizophrenia, living in five European countries, was explored using well-validated questionnaires. The adoption of problem-focused coping strategies was more frequent among young relatives and among relatives of younger patients, and was associated with higher levels of practical and emotional social support and of professional help. In contrast, emotion-focused strategies were more frequently adopted by relatives who had been living longer with the patient and who had poorer social support. It is suggested that supportive and educational interventions should be provided as early as possible to relatives of patients with schizophrenia, which, in addition to having a practical focus, should also have a social focus, aiming at extending the family's social network.

In vivo efficacies of 5'-methylthioadenosine analogs as trypanocides.

5'-Deoxy-5'-(methylthio)adenosine (MTA), a key by-product of polyamine biosynthesis, is cleaved by MTA phosphorylase and is salvaged as adenine and, through conversion of the ribose moiety, methionine. An analog of MTA, 5'-deoxy-5'-(hydroxyethylthio)adenosine (HETA), is a substrate for trypanosome MTA phosphorylase and is active in vitro and in vivo against Trypanosoma brucei brucei, an agent of bovine trypanosomiasis. In this study, HETA and three O-acylated HETA derivatives were examined for their activities against model infections of T. b. brucei and Trypanosoma brucei rhodesiense, the agent of East African sleeping sickness. HETA was curative (>60%) for infections caused by 5 of 11 clinical isolates of T. b. rhodesiense when it was given to mice at 200 mg/kg of body weight for 7 days as a continuous infusion in osmotic pumps. HETA at 150 to 200 mg/kg also extended the life spans of the mice infected with four additional isolates two- to fivefold. Di- and tri-O-acetylated derivatives of HETA also proved curative for the infections, while a tri-O-propionyl derivative, although also curative, was not as effective. This study indicates that substrate analogs of MTA should be given important consideration for development as novel chemotherapies against African trypanosomiasis.

Antitrypanosomal activity of purine nucleosides can be enhanced by their conversion to O-acetylated derivatives.

Fifteen purine nucleosides and their O-acetylated ester derivatives were examined for in vitro antitrypanosomal activity against the LAB 110 EATRO isolate of Trypanosoma brucei brucei and two clinical isolates of Trypanosoma brucei rhodesiense. Initial comparisons of activity were made for the LAB 110 EATRO isolate. Three nucleoside analogs exhibited no significant activity (50% inhibitory concentrations [IC50s] of > 100 microM), whether they were O acetylated or unacetylated; three nucleosides showed almost equal activity (IC50s of < 5 microM) for the parent compound and the O-acetylated derivative; nine nucleosides showed significantly improved activity (> or = 3-fold) upon O acetylation; of these nine analogs, six displayed activity at least 10-fold greater than that of their parent nucleosides. The most significant results were those for four apparently inactive compounds which, upon O acetylation, displayed IC50s of < or = 25 microM. When the series of compounds was tested against T. brucei rhodesiense isolates (KETRI 243 and KETRI 269), their antitrypanosomal effects were comparable to those observed for the EATRO 110 strain. Thus, our studies of purine nucleosides have determined that O acetylation consistently improved their in vitro antitrypanosomal activity. This observed phenomenon was independent of their cellular enzyme targets (i.e., S-adenosylmethionine, polyamine, or purine salvage pathways). On the basis of our results, the routine preparation of O-acetylated purine nucleosides for in vitro screening of antitrypanosomal activity is recommended, since O acetylation transformed several inactive nucleosides into compounds with significant activity, presumably by improving uptake characteristics. O-acetylated purine nucleosides may offer in vivo therapeutic advantages compared with their parent nucleosides, and this possibility should be considered in future evaluations of this structural class of trypanocides.

A new questionnaire assessing coping strategies in relatives of patients with schizophrenia: development and factor analysis.

This paper describes the development and validation of a questionnaire assessing the coping strategies adopted by relatives of patients with schizophrenia. The final version of the questionnaire includes 27 items, grouped into seven subscales (information, positive communication, social interests, coercion, avoidance, resignation and patient's social involvement), the intra-rater reliability of which ranges from 0.46 to 0.76. Cronbach's alpha coefficient, which tests the content validity of the subscales, ranges from 0.68 to 0.83. Factor analysis identifies three factors (problem-oriented coping strategies, emotionally focused strategies, and maintenance of social interests in association with patient's avoidance), accounting for 70.9% of the total variance. This questionnaire may be particularly useful for targeting and monitoring psychoeducational interventions in the families of patients with schizophrenia.

Burden on the families of patients with obsessive-compulsive disorder: a pilot study.

This study aimed to explore the burden on the key-relatives of a sample of patients with obsessive-compulsive disorder and a control sample of patients with major depression. The level of burden of relatives of obsessive-compulsive patients was moderate to severe in most of the explored situations, and did not differ significantly from that of the relatives of depressives. Social relationships were compromised in 74% of cases, and feelings of depression were recorded in 84% of cases. The level of subjective burden changed according to the relatives' sex, age and position in the family. The degree of objective burden correlated with the level of patients' disability, whereas that of subjective burden correlated with the severity of patients' obsessive-compulsive symptomatology.

5-Methyl-2'-deoxycytidine in single-stranded DNA can act in cis to signal de novo DNA methylation.

Methylation of cytosine residues in DNA plays an important role in regulating gene expression during vertebrate embryonic development. Conversely, disruption of normal patterns of methylation is common in tumors and occurs early in progression of some human cancers. In vertebrates, it appears that the same DNA methyltransferase maintains preexisting patterns of methylation during DNA replication and carries out de novo methylation to create new methylation patterns. There are several indications that inherent signals in DNA structure can act in vivo to initiate or block de novo methylation in adjacent DNA regions. To identify sequences capable of enhancing de novo methylation of DNA in vitro, we designed a series of oligodeoxyribonucleotide substrates with substrate cytosine residues in different sequence contexts. We obtained evidence that some 5-methylcytosine residues in these single-stranded DNAs can stimulate de novo methylation of adjacent sites by murine DNA 5-cytosine methyltransferase as effectively as 5-methylcytosine residues in double-stranded DNA stimulate maintenance methylation. This suggests that double-stranded DNA may not be the primary natural substrate for de novo methylation and that looped single-stranded structures formed during the normal course of DNA replication or repair serve as "nucleation" sites for de novo methylation of adjacent DNA regions.

Clinical and socio-demographic correlates of coping strategies in relatives of schizophrenic patients.

The study aimed to describe coping strategies in a sample of key-relatives of schizophrenic patients, and to explore their relationships with the following variables: patients' and key-relatives' socio-demographic characteristics; patients' and key-relatives' clinical status; and key-relatives' subjective burden. Significant correlations were found between the scores on coercion items and those on Present State Examination (PSE-9) factors "disorganized syndrome" and "psychomotor poverty syndrome". A long duration of illness was found to be associated with relatives' strategies characterized by positive communication and patient's social involvement. Coercion, avoidance and resignation coping strategies were significantly associated with the occurrence of anxiety and depressive symptoms in patients' key-relatives.