Clinical profile of schizophrenia comorbid with obsessive-compulsive symptoms: A comparative study.

OBJECTIVE: Describe and compare the clinical profile of schizophrenic patients with and without obsessive-compulsive symptoms and obsessive-compulsive disorder (OCD) patients. METHODS: A descriptive and analytical cross-sectional study was carried out at the psychiatry department of Hassan II University Hospital in Fez over 12 months to compare three groups of patients: "schizo-obsessive" (n=32), "schizophrenia" (n=34), and "OCD" (n=46). All participants (n=112) were assessed using the mini-international neuropsychiatric interview (MINI), the Yale-Brown obsessive-compulsive scale (Y-BOCS), the Brown assessment of beliefs scale (BABS), the Hamilton anxiety rating scale (HAM-A), the Beck's depression inventory (BDI-II), the positive and negative syndrome scale (PANSS), and the clinical global impressions-severity scale (CGI-S). RESULTS: The "schizo-obsessive" group differed from the "schizophrenia" group in: more severe psychotic symptoms (mean=64.16±17.049, P<0.001), higher anxiety (mean=8.87±5,655, P<0.001) and depression (mean=7.50±5.989, P<0.001) scores, more prevalent suicide attempts (46.9%), higher illness severity score (mean=5.13±1.157, P=0.02), and more professional disinsertion (78.1%). The "schizo-obsessive" group (mean= 14.47±3.388) had significantly poor insight (P<0.001) compared to the "OCD" group (mean= 8.35±4.542). There were similarities in the obsessive and compulsive themes between the "schizo-obsessive" and the "OCD" groups, with no significant difference of severity (P=0.26). CONCLUSION: A careful assessment of obsessive symptomatology is essential in schizophrenia for better patient management and prognosis.

Stigmatizing attitudes and social perception towards mental illness among Moroccan medical students.

OBJECTIVES: This study aimed to explore medical students' attitudes, social perception, and knowledge towards mental illness and identify the associated factors. METHODS: A cross-sectional study was conducted in the Medical Faculty of Fez. A representative sample of 420 Moroccan students from the first to the seventh years was selected randomly during the academic year (2018/2019). They completed the Mental Illness Clinicians Attitudes scale (MICA) and a self-questionnaire exploring sociodemographic data, the university course, social perception and knowledge towards mental illness, and the potential consideration of psychiatry as a career. RESULTS: The average age of participants was 21.73 years (SD ±2.60), and 72.4% (n=304) of respondents were female (M/F gender ratio=0.39). The participants held stigmatizing attitudes and negative social perceptions towards mental illness, as indicated by the high score of the MICA scale (mean=57.24; SD ±9.95). Females (p=0.02) and married students (p=0.02) showed significantly more tolerant attitudes. The attitudes of students (p=0.37) who completed the psychiatry clerkship were slightly more favorable (p=0.15). There was no significant difference in attitudes according to the level of study (p=0.06). Students with a lower socioeconomic level tended to be less stigmatizing (p=0.08). The assessment of knowledge about mental illness among students objectified major gaps. A total of 17.9% (n=75) considered psychiatry as a career. CONCLUSION: The attitudes of medical students in this study were stigmatizing, and this should without delay motivate Moroccan research, educational and health authorities to investigate further scientific research in this area to address these attitudes and remedies.

Cultural Adaptation and Validation of the Modified Checklist for Autism in Toddlers, Revised with Follow-up in Moroccan Arabic dialect.

BACKGROUND: The prevalence of autism spectrum disorder is increasing worldwide, making screening and early intervention necessary. Several screening instruments have been developed in recent years. The Modified Checklist for Autism in Toddlers Revised with Follow-up (M-CHAT-R/F) is considered to be one of the specific measures designed to identify toddlers at risk for autistic spectrum disorder. OBJECTIVE: The aim of the study was to translate and adapt the original version of M-CHAT-R/F from the English to the Moroccan Arabic language. STUDY DESIGN: Specialized translators and clinicians ensured forward and backward translation of the scale into Moroccan Arabic. Then, a two-stage screening of the M-CHAT-R/F-T was applied to a study sample comprised of 56 toddlers with autistic spectrum disorder (category I) and 96 toddlers with normal development (category II). "Kappa test", "Cronbach's alpha" test, the intra class correlation coefficient, and the area under the curve were determined. RESULT: The average score results of M-CHAT-R/F were 13.12 for category I, while it was 2.24 for category II. The Cronbach's alpha coefficient of the checklist was 0.929. The kappa values ranged from k=0.78 to k=0.97 with a confidence interval of 95% indicating good convergence. The intra-class correlation coefficient ranged from 0.97 to 0.99, which is excellent. The area under the curve in our study was 0.988, an excellent result. CONCLUSION: Efficiency of the Moroccan Arabic version of the MCHAT was demonstrated for screening in the general population.

[Clinical presentation, co-occurrence and suicidality in psychiatric emergency patients with psychogenic non-epileptic seizures]. / Expression clinique, comorbidités psychiatriques et suicidalité chez des patients avec crises psychogènes non épileptiques.

INTRODUCTION: Non-epileptic psychogenic seizures (PNES) are a common condition but little studied in psychiatry. Our study is the first Moroccan of this topic. Our aim was to describe their clinical presentation, comorbidities and suicidality. METHOD: It was a one-year recruitment study conducted in the Department of Psychiatry at the University Hospital of Fez. We collected socio-demographic data. Psychometric evaluation used the MINI criteria for different psychiatric disorders, personality disorders and suicidality, the Hamilton anxiety scale, the Beck depression scale and the Childhood Trauma Questionnaire. RESULTS: Thirty-two patients were included, the majority of whom were female with a low economic level (89 %). The average age of onset average was 23.97 years. The generalized form represented 71.9 % of cases with a triggering factor present in 46 %. The duration of the seizures average was 23.47minutes, and 37.5 % of the patients had at least one attack per day. Among patients, 68.8 % had comorbidity: major depressive disorder MDD (46 %) and post-traumatic stress disorder PTSD (18.6 %). Suicide risk was rated as moderate or severe in 12.5 % of patients, and 40.62 % had a previous suicide attempt. A personality disorder was present in 18.8 % of cases. Finally, a history of child abuse was found in 59.4 % of patients and was correlated to PNES age of onset and co-occurrence depression. CONCLUSION: We found several similarities with previous studies in terms of clinical description and co-occurrences. However, our sample is characterized by more prolonged and more frequent seizures and a higher rate of suicidality.

[The role of the general practitioner in management of psychiatric disorders]. / Le rôle du médecin généraliste dans la prise en charge des pathologies psychiatriques.

AIM: Psychiatric disorders represent a huge burden of disease worldwide. Significant gaps in their diagnosis and treatment remain. In Morocco, an observed shortage of psychiatrists would logically call for their collaboration with family physicians. Our objectives were to analyse perceptions of general practitioners' capacity to manage psychiatric disorders, to study the difficulties encountered in their management of these pathologies, and to assess their degree of collaboration with existing mental health structures. METHODS: A cross-sectional nationwide survey of family physicians practicing in urban and rural areas in the private and public sectors in the Kingdom of Morocco was organised. Carried out over a period of 10 months (April 2016 to January 2017), the study was based on an anonymised self-administered questionnaire comprising 23 items. Were excluded from the sample: physicians who had tested the questionnaire prior to its distribution, medical students, family physicians employed in administrative services, resident doctors in university hospitals, and questionnaires with incomplete answers. RESULTS: All in all, five hundred and four questionnaires were distributed, with a participation rate of 79.8%, while 57.9% of the respondents were men. Their seniority as doctors ranged from 1 year to 60 years with average duration of 19.5±11 years. An overwhelming majority of the doctors had obtained their doctorates in Morocco, while 8.4% had been trained abroad. The majority of the doctors surveyed worked in private practice in urban areas (53.1%), and most of them (81.9%) agreed that the management of psychiatric disorders is a public health priority. The participating physicians stated that they received an average of five patients per day suffering from mental disorders, representing 17.3% of their medical consultations. According to their perceptions, the most frequent psychiatric pathologies were depression, somatoform disorders and sleep disturbances, which were cited by 92.9%, 78.8% and 78.5% of them respectively. Regarding the diagnosis of psychiatric pathologies, 60.2% of the doctors considered bipolar disorder as a difficult pathology to diagnose, followed by schizophrenia, autism and dementia, while the pathologies reported as the most difficult to treat were addictive behaviours, schizophrenia, bipolar disorder, autism and dementia. Physicians felt a need for training, primarily pertaining to anxiety disorders, schizophrenia and bipolar disorders (49.5%, 35% and 33.7%, respectively). Nearly a quarter of the respondents (22.4%) mentioned a need for training in management of all psychiatric illnesses. Regarding the reasons for their referral of patients to mental health care structures, 65.4% of the physicians justified their doing so by a need for hospitalisation, while in 43.7% of cases, the transfer was carried out at the request of the patient or his family. The difficulties mentioned by the respondents were firstly a lack of adapted means of care (77.5%), followed by their insufficient training in mental healthcare (52.4%) and absence of collaboration with mental health professionals (52.4%). CONCLUSION: For the majority of the physicians surveyed, psychiatry represented a public health priority, and their prioritising provides a probable explanation for their positive perception of their role in the management of mental disorders. However, we went on to observe that a number of difficulties constituted obstacles to their role in management of mental disorders and induced them to refer their patients to psychiatrists. Our analysis highlighted the impact of continuing education on physicians' attitudes and the interest of collaboration between practitioners. Efforts in university teaching and continuous education should be encouraged in view of enabling family physicians to more effectively cope with the demands of daily practice and meet the expectations of the overall population.

Psychoactive substances use and associated factors among middle and high school students in the North Center of Morocco: a cross-sectional questionnaire survey.

BACKGROUND: Data on psychoactive substance (PAS) consumption among adolescents in the North Center of Morocco are not at all available. Therefore, the current study aimed at investigating the prevalence and the determinants of psychoactive substances use among middle and high school students in this region. METHODS: A cross-sectional study was conducted from April 2012 to November 2013 in public middle and high schools in the North Central Region of Morocco. An anonymous self-administered questionnaire was used to assess psychoactive substances use among a representative sample of school students from the 7th to the 12th grade, aged 11-23 years, selected by stratified cluster random sampling. Factors associated with psychoactive substance use were identified using multivariate stepwise logistic regression analyses. RESULTS: A total of 3020 school students completed the questionnaires, 53.0 % of which were males. The overall lifetime smoking prevalence was 16.1 %. The lifetime, annual and past month rates of any psychoactive substance use among the study subjects were 9.3, 7.5, and 6.3 % respectively. Cannabis recorded the highest lifetime prevalence of 8.1 %, followed by alcohol 4.3 %, inhalants 1.7 %, psychotropic substances without medical prescription 1.0, cocaine 0.7, heroine 0.3, and amphetamine with only 0.2 %. Psychoactive substance use was associated with males more than females. The risk factors identified by multivariate stepwise logistic regression analyses were being male, studying in secondary school level, smoking tobacco, living with a family member who uses tobacco, and feeling insecure within the family. CONCLUSIONS: The prevalence among all school students reported by the current study was comparable to the national prevalence. Efforts to initiate psychoactive substance prevention programs among school students should be made by designing such programs based on the significant factors associated with psychoactive substance use identified in this study.

[Reversible catatonia after the abrupt discontinuation of clozapine: Case report]. / Une catatonie réversible suite à l'arrêt brutal de la clozapine. À propos d'un cas.

In this paper, we report the case of a patient, aged 26, with schizophrenia who was admitted to psychiatric emergencies for catatonia, one week after abrupt discontinuation of clozapine. An improvement was seen after only two days of the reintroduction of clozapine alone. This catatonia is reversible and it responds magnificently to the reintroduction of clozapine. And we conclude that patients and their caregivers need to be educated about the effects of abrupt cessation of clozapine administration.

Suicidal behaviors among Moroccan school students: prevalence and association with socio-demographic characteristics and psychoactive substances use: a cross-sectional study.

BACKGROUND: Suicidal behavior is a major cause of injury and death worldwide, especially among adolescents and young adults. Few studies have tackled this issue in the Arab world. The present study investigated the prevalence and the risk factors of suicidal behaviors among Moroccan school students. METHODS: From April 2012 to November 2013, a cross-sectional study was conducted in the North-Centre region of Morocco among students in public secondary schools selected using stratified cluster random sampling. The data were collected via anonymous self-administered questionnaires. The Mini International Neuropsychiatric Interview was used in its Moroccan Colloquial Arabic version to assess suicidality according to the DSM-IV criteria. RESULTS: A total of 3020 students (53 % boys) aged 11-23 years (average age = 16 ± 2.1 years) were included in the study. The prevalence of suicide ideation, suicide planning and suicide attempts during the last month were 15.7, 6.3, and 6.5 % respectively. Univariate analyses demonstrated that suicidal behaviors followed different epidemiological patterns. According to the multivariate analyses, the risk factors for all suicidal behaviors among Moroccan school students were the female gender, middle school level, urban locations, low family income, parents' divorce, tobacco consumption and psychoactive substances (alcohol and cannabis) use. CONCLUSIONS: The intervention of preventive programs has become an emergency to overcome the issue of suicidality in Morocco. Further researches on adolescents' suicidal behaviors are suggested to update temporal data and assess the effectiveness of potential interventions.

Treatment of malignant gliomas with a replicating adenoviral vector expressing herpes simplex virus-thymidine kinase.

We evaluated the interaction between oncolytic, replication-competent adenoviral vectors and the herpes simplex virus-1 thymidine kinase (HSV1-tk) gene/ganciclovir (GCV) suicide system for the treatment of malignant gliomas. We constructed a panel of replication-competent adenoviral vectors in which the luciferase (IG.Ad5E1(+). E3Luc) or HSV1-tk gene (IG.Ad5E1(+).E3TK) replace the M(r) 19,000 glycoprotein (gp19K) coding sequence in the E3 region. IG.Ad5E1. IG.Ad5.ClipLuc and IG.AdApt.TK are E1-deleted viruses that contain the luciferase or the HSV1-tk gene in the former E1 region driven by the human cytomegalovirus promoter. IG.Ad5. Sarcoma 1800HSA.E3Luc contains an irrelevant gene in the E1 region, whereas the gp19K coding sequence in the E3 region is replaced by the luciferase gene as in the replicating virus IG.Ad5E1(+).E3Luc. For in vitro experiments, we used a panel of human glioma cell lines (U87 MG, T98G, A172, LW5, and U251), a rat gliosarcoma cell line (9 L), and human lung (A549) and prostate carcinoma (P3) cell lines. In vitro, GCV sensitivity (10 microg/ml) was studied in U87 MG cells after infection at a multiplicity of infection of 1 and 10. A s.c. U87 MG glioma xenograft model was established in NIH-bg-nu-xid mice. Tumors of 100-150 mm(3) were treated with a single injection of adenovirus 10(9) IU suspended in 100 microl of PBS, and GCV 100 mg/kg was administered i.p. twice daily for 7 days. The cytopathic effect of all three replication-competent adenoviral vectors was similar to the cytopathic effect of wild-type adenovirus 5 on all human cell lines tested, indicating that deletion of the E3 gp19K sequences did not affect the oncolytic effect of the vectors. In vitro, luciferase expression was the same for both E1-deleted vectors (IG.Ad5.ClipLuc and IG.Ad5. Sarcoma 1800HSA.E3Luc), demonstrating the strength of the internal E3 promoter even in the absence of E1A. However, in vitro expression levels obtained with replication-competent IG.Ad5E1(+). E3Luc were 3 log higher (allowing infection with a 2-3-log lower multiplicity of infection) in the human cell lines. In U87 MG glioma cells, the oncolytic effect of replication-competent IG.Ad5E1(+).E3TK was significantly enhanced by the addition of GCV and greatly exceeded the cytotoxicity of replication-incompetent IG.AdApt.TK combined with GCV. In established s.c. U87 MG glioma xenografts, a single injection of IG.Ad5E1(+).E3TK resulted in a significant slowing of tumor growth and prolonged survival compared with injection of IG.AdApt.TK. Addition of GCV slowed tumor growth, further adding to survival. In conclusion, the oncolytic effect of replicating adenoviral vectors and HSV1-tk/GCV have potent antitumor effects in gliomas. When combined, these two approaches are complementary, resulting in a significantly improved treatment outcome. In addition, replication-competent adenoviral vectors missing the E3 gp19K coding sequences, have oncolytic efficacy comparable with wild type. In combination with high expression levels obtained with the natural E3 promoter, such vectors are promising new anticancer agents.

In vitro and in vivo gene transfer to pulmonary cells mediated by cationic liposomes.

Cationic liposomes have been proposed as alternative to adenovirus in the treatment of cystic fibrosis lung disease. Therefore, we have investigated the efficiency of two lipid mixtures in mediating gene transfer in in vitro and in vivo models. The cationic lipid DOTMA (N-(1-(2,3(dioleyloxy)propyl)-n,n,n-trimethylammoniumchloride++ +) and DOGS (dioctadecylamidoglycylspermine) were used in combination with the neutral lipid DOPE (dioleoylphosphatidylethanolamine). The relative transfection efficiencies of the two cationic liposomes were tested using the bacterial beta-galactosidase (lacZ) and the firefly luciferase genes. Gene expression was detected in both cell limes and primary culture of rhesus monkey airway epithelium after transfection with plasmid DNA complexed with DOGS/DOPE or DOTMA/DOPE. Transfection efficiency of both types of lipids was higher in the mouse fibroblast 3T3 cell line as compared to human carcinoma A549 cells and primary epithelial cultures. Administration of DNA-liposome complexes via intratracheal instillation resulted in expression of the lacZ and luciferase marker gene in the mouse airways. In vivo transfection mediated by both types of liposomes were proven to be far less efficient than adenovirus treatment.

Characterization of the gene encoding human peroxisomal 3-oxoacyl-CoA thiolase (ACAA). No large DNA rearrangement in a thiolase-deficient patient.

We have characterized the gene encoding human peroxisomal 3-oxoacyl-CoA thiolase, an enzyme operative in the peroxisomal beta-oxidation system. We found one version of this gene (gene symbol ACAA) in the human genome, in contrast to the situation in rat where two versions have been described. The human gene shows a high structural similarity to the rat genes. It contains 12 exons and 11 introns and spans about 11 kb. We have determined the 5' end of the human thiolase mRNA by employing primer extension analysis and we have sequenced the region upstream of the gene. The putative promoter area displays some of the characteristics typical of promoters of other peroxisomal genes, in that it contains GC elements, but lacks TATA boxes. Finally, no large DNA rearrangement involving the thiolase gene could be observed in a patient suffering from pseudo-Zellweger syndrome (peroxisomal thiolase deficiency).

Zonal distribution of peroxisomal 3-oxoacyl-CoA thiolase mRNA in liver from rats treated with di-(2-ethylhexyl)phthalate.

Treatment of rats with di-(2-ethylhexyl)phthalate leads to a dramatic increase in peroxisomal 3-oxoacyl-CoA thiolase RNA, the concentration being higher in the pericentral than in periportal hepatocytes. These findings indicate that the production of peroxisomal thiolase and the zonal distribution of the enzyme are regulated at a pretranslational level.

Lung gene therapy: in vivo adenovirus-mediated gene transfer to rhesus monkey airway epithelium.

Somatic gene therapy of lung disorders such as cystic fibrosis (CF) aims at introducing the therapeutic gene into respiratory epithelium. We have tested the ability of recombinant human adenovirus to infect rhesus monkey airway epithelium in vivo. Application of adenovirus harboring the lacZ marker gene to the airway surface resulted in large patches of lacZ-positive cells in the trachea, bronchi, and bronchioles, 6 days after virus exposure, indicating a successful transfer of the lacZ gene to respiratory epithelium. Microscopic analysis showed that basal, mucous goblet, and ciliated cells were lacZ positive. In addition, gene transfer to the submucosal glands was observed. Pathological examination of the organs revealed no virus-mediated toxic effects to the lungs and other organs. Using polymerase chain reaction (PCR) analysis we found no spread of the virus to blood or any organ tested. These results indicate the potential use and safety of adenoviruses as a tool in human gene therapy procedures aimed at pulmonary diseases.

Feasibility of adenovirus-mediated nonsurgical synovectomy in collagen-induced arthritis-affected rhesus monkeys.

Gene transfer to synovial tissue by adenoviral vectors (Ad) was studied in vitro in cultured human synoviocytes and in vivo in seven primates with arthritis. Hyperplastic synovium was efficiently transduced with Ad.lacZ in vitro and in vivo in rhesus monkeys with collagen-induced arthritis, whereas chondrocytes were not transduced. Intraarticular injection of recombinant Ad harboring the luciferase gene showed the presence of reporter gene products only in Ad-injected joints. In addition, the feasibility of synovectomy by Ad harboring the herpes simplex virus thymidine kinase gene (tk) was studied. In vitro infection of synovium from rheumatoid arthritis patients with Ad.TK, followed by administration of ganciclovir, resulted in death of >90% of the synoviocytes. By mixing Ad.TK-infected with noninfected cells, it appeared that the presence of 10% infected synoviocytes resulted in the killing of more than 85% of the synoviocytes, demonstrating a substantial bystander effect. Intraarticular injection of Ad.TK in the knees of rhesus monkeys with arthritis, followed by treatment with ganciclovir for 14 days, resulted in increased apoptotic cell death in the synovium of Ad.TK-injected as compared with noninjected joints and ablation of the synovial lining layer. The procedure revealed no toxic side effects. These data suggest that nonsurgical synovectomy by tK gene therapy is feasible.

Aerosol-mediated delivery of recombinant adenovirus to the airways of nonhuman primates.

At present, it is conceivable that gene therapy of the cystic fibrosis airway epithelium is possible using the direct transfer of a functional human cystic fibrosis transmembrane conductance regulator (CFTR) gene to a wide variety of patients' tracheo-bronchial cells. Here we describe a novel approach (aerosolization) to deliver a replication-deficient adenovirus carrying the CFTR gene (Ad.CFTR) to the airways. Results obtained in vitro and in Rhesus monkeys suggest that the delivery of recombinant adenovirus as an aerosol is feasible and is not associated with severe toxicity after single or double administration depending on the Ad.CFTR dose. This study supports the concept of aerosolization as a delivery method for adenovirus-mediated lung gene therapy.

Trans-complementation of E1-deleted adenovirus: a new vector to reduce the possibility of codissemination of wild-type and recombinant adenoviruses.

Treatment of cystic fibrosis by gene therapy will require the development of vectors capable of efficient and safe transfer of a functional cystic fibrosis transmembrane conductance regulator (CFTR) cDNA to airway epithelia. To achieve this goal, replication-deficient (E1-) adenoviruses (Ad) are promising vectors. We have previously demonstrated efficient CFTR gene delivery to the airways of cotton rats and rhesus monkeys using a replication-deficient adenovirus, Ad-CFTR. Here, we have investigated an important safety issue, the interaction between the vector and wild-type virus which can provide the missing E1 function in trans. We show that Ad5 can mobilize the defective Ad-CFTR genome in vitro and in cotton rats. However, the extent of the complementation in vivo by wild-type virus is limited because no additional spreading or shedding of Ad-CFTR to trachea, lungs, and stools is elicited. To attenuate Ad-CFTR further, a mutation was introduced in the cis-acting regulatory sequences that control the encapsidation of the viral genome. We demonstrate that when cells are coinfected with wild-type virus and the new attenuated vector, the viral DNA containing the natural encapsidation sequences is preferentially packaged, leading to a rapid dilution of the recombinant virus.

Distribution of recombinant adenovirus in the cerebrospinal fluid of nonhuman primates.

Gene therapy by administration of vectors into the cerebrospinal fluid (CSF) may be used in treatment of leptomeningeal metastases (cancer gene therapy) as well as in treatment of neurodegenerative disorders, traumatic injury, and chronic pain. Recombinant adenoviruses are attractive vectors for intra-CSF administration because they can efficiently transfer genes into the nonreplicating cells of the central nervous system (CNS). In addition, they can be produced in high titers and, because no producers cells are introduced, the risk of CSF obstruction by clustering cells is circumvented. However, successful application requires favorable distribution dynamics, high transduction efficiency, and long-lasting transgene expression. In this study we examined the distribution of a recombinant adenovirus containing the lacZ gene after administration into the CSF of nonhuman primates. After intraventricular and suboccipital administration, homogeneous distribution of the vector along the meninges covering the brain and spinal cord was obtained, as demonstrated by extensive and intense blue staining of cells, predominantly in the arachnoid and pia mater. In one animal we also found beta-galactosidase activity in the cervical paraspinal fat and in one of the deep cervical lymph nodes, indicating drainage of the vector or vector products with CSF into cervical lymph. This route of vector clearance from the CNS may result in antigenic presentation and an effective immune response and may explain the sixfold higher serum antibody titers after intrathecal injection of adenovirus as compared with intranasal application in Fischer rats. We conclude that distribution dynamics of recombinant adenovirus after intra-CSF administration are excellent. However, because of the immune response elicited by the virus, even after administration to the CNS, development of immunomodulating strategies remains a challenge.

Adenovirus-mediated interleukin 3 beta gene transfer by isolated limb perfusion inhibits growth of limb sarcoma in rats.

Cytokine gene transfer using (multiple) intratumoral injections can induce tumor regression in several animal models, but this administration technique limits the use for human gene therapy. In the present studies we describe tumor growth inhibition of established limb sarcomas after a single isolated limb perfusion (ILP) with recombinant adenoviral vectors harboring the rat IL-3 beta gene (IG.Ad.CMV.rIL-3 beta). In contrast, a single intratumoral injection or intravenous administration did not affect tumor growth. Dose-finding studies demonstrated a dose-dependent response with a loss of antitumor effect below 1 x 10(9) IU of IG.Ad.CMV.rIL-3 beta. Perfusions with adenoviral vectors bearing a weaker promoter (MLP promoter) driving the rIL-3 beta gene did not result in antitumor responses, suggesting that the rIL-3 beta-mediated antitumor effect depends on the amount of rIL-3 beta protein expressed by the infected cells. Furthermore, it was shown by direct comparison that ILP with IG.Ad.CMV.rIL-3 beta in the ROS-1 osteosarcoma model is at least as efficient as the established therapy with the combination of TNF-alpha and melphalan. Treatment with IG.Ad.CMV.rIL-3 beta induced a transient dose-dependent leukocytosis accompanied by an increase in peripheral blood levels of histamine. Leukocyte infiltrations were also histopathologically demonstrated in tumors after perfusion. These results demonstrate that ILP with recombinant adenoviral vectors carrying the IL-3 beta transgene inhibits tumor growth in rats and suggest that cytokine gene therapy using this administration technique might be beneficial for clinical cancer treatment.