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Italian thistle (Carduus pycnocephalus L.) is a common and increasingly important weed in Tunisia. It is also problematic in the western United States and a target of biological control. In surveys conducted in northern Tunisia from 2003 to 2005, Italian thistle plants in many locations were found diseased by rust. Eighty-five isolates of rust were collected from Italian thistle during these surveys. Each isolate was collected from a single plant and stored individually as mixtures of urediniospores and teliospores at 4°C or in liquid nitrogen. Urediniospores and teliospores of all isolates were similar in morphology and matched the description of Puccinia carduorum Jacky (3). Isolate B1003 (BPI No. 878207), collected from Béja, Tunisia, was arbitrarily selected for further study. Comparison of internal transcribed spacer (ITS) regions of B1003 (GenBank Accession No. EF050059) with other ITS sequences indicated a 97% similarity to P. carduorum (GenBank Accession No. PCU57351) from Carduus nutans subsp. leiophyllus. Eight Italian thistle plants, grown from seeds collected in Béja, were inoculated in the 3- to 5-leaf stage with urediniospores of B1003 by spraying plants with an aqueous suspension of urediniospores at 106 spores per ml with approximately 0.03 ml of surfactant until they were thoroughly wet. Four plants were sprayed with water plus surfactant only. All plants were covered with plastic bags and placed in a growth chamber at 18/20°C night/day temperatures. Bags were removed 24 h after inoculation and plants were monitored daily for symptoms. Plants sprayed with water plus surfactant only did not develop symptoms. Six inoculated plants developed disease symptoms similar to those observed on samples collected during the surveys. White flecks appeared within 7 days of inoculation and developed into brown pustules 10 days after inoculation. Pustules enlarged and produced urediniospores until they covered both sides of diseased leaves. One month after appearance of symptoms, diseased leaves turned yellow and died. Urediniospores from these plants were used to inoculate six plants each at the 2- to 5-, 6- to 8-, and >8-leaf stages. All plants became diseased and produced uredinia. Plants in the 2- to 5-leaf stage were more severely diseased than other plants. P. carduorum was introduced to the United States in a field test for control of C. nutans subsp. leiophyllus (musk thistle) and has become established in a number of states (1). An isolate of this fungus has also been found from C. tenuiflorus (slender-flower thistle) in California (4). However, neither isolate causes substantial disease on Italian thistle from California (2,4). Isolates of P. carduorum from C. pycnocephalus in Greece, Italy, and Turkey caused little disease on most Italian thistle collections from California (2), indicating variability in susceptibility among plants as well as in virulence among rust isolates from different geographical locations. To our knowledge, this is the first report of P. carduorum parasitizing Italian thistle in Tunisia. Tests will be conducted to determine the host range of this isolate among C. pycnocephalus collections and other species. References: (1) A. B. A. M. Baudoin and W. L. Bruckart. Plant Dis. 80:1193, 1996. (2) W. L. Bruckart and G. L. Peterson. Phytopathology 81:192, 1991. (3) D. B. O. Savile. Can. J. Bot. 48:1553, 1970. (4) A. K. Watson and K. Brunetti. Plant Dis. 68:1003, 1984.

Emex spinosa (L.) is a common weed in cereal crops and pastures in northern Tunisia. The build up of the seed bank from a cropping-grazing farming system makes chemical and cultural controls inefficient. Biological control as part of integrated weed management may improve weed control. Diseased seedlings were collected from several locations in northern Tunisia during field surveys. Symptoms were small, circular, light brown leaf spots varying in size (1.75 to 3.5 mm in diameter) with a definite dark brown border on both sides of leaves that wilted and died. Microscopic observations showed conidiophores and conidia within and around the spots. Pure cultures from single conidia were obtained on carrot leaf extract agar. The fungus was identified as Cercospora tripolitana on the basis of identification keys described by Chupp (1). Conidia, borne on unbranched, fasciculated conidiophores, were elongate, hyaline, multiseptate, 110 to 150 × 1.8 to 3.7 μm (average 130 to 2.5 μm), and had truncate bases. For pathogenicity testing, six plants were sprayed with a spore suspension of 6 × 105 conidia/ml. Controls were sprayed with sterile distilled water. Plants were placed in a growth chamber at 22°C, 95% relative humidity, and 18/6 h of light/dark and monitored for symptoms. Ten days after inoculation, symptoms identical to those observed in the field were observed on inoculated plants. Control plants did not develop any symptoms. Four weeks later, diseased leaves turned yellow and died. The fungus was reisolated from symptomatic plants according to Koch's postulates. Although C. tripolitana has been previously reported in North and South Africa, to our knowledge, this is the first report of the fungus as a pathogen on E. spinosa under Tunisian agroecological conditions, making it a promising candidate for weed control. Reference: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. C. Chupp, Ithaca, New York, 1953.

The herbaceous perennial Lepidium draba L. is an invasive weed of rangelands and riparian areas in North America and Australia. As of 2002, it had infested 40,500 ha of rangeland in Oregon and large areas in Wyoming and Utah. Little is known of plant pathogens occurring on L. draba, especially in the United States, that could be useful for biological control of the weed. Leaf spots were first noted on a stand of L. draba near Shepherd, MT in 1997. The spots were mostly circular but sometimes irregularly shaped and whitish to pale yellow. The pathogen was erroneously assumed to be Cercospora beticola since its morphological traits closely resembled that species and the area had large fields of sugar beet with heavy Cercospora leaf spot incidence. Diseased leaves of L. draba were collected in 1997 and 2007. Conidia, borne singly on dark gray, unbranched conidiophores produced on dark stromata late in the season, were elongate, hyaline, multiseptate, 38 to 120 × 2 to 6 μm (mostly 38 to 50 × 2 to 5 μm) and had bluntly rounded tips and wider, truncate bases. These characteristics were consistent with the description of C. bizzozeriana Saccardo & Berlese (2). To isolate the fungus, spores were picked from fascicles of conidiophores with a fine-tipped glass rod, suspended in sterile water, and spread on plates of water agar. Germinated spores were transferred to potato dextrose agar (PDA). The ITS1, 5.8S, and ITS2 sequences of this fungus (GenBank Accession No. EU887131) were identical to sequences of an isolate of C. bizzozeriana from Tunisia (GenBank Accession No. DQ370428). However, these sequences were also identical to those of a number of Cercospora spp. in GenBank, including C. beticola. We also compared the actin gene sequences of the Montana isolate of C. bizzozeriana (GenBank Accession No. FJ205397) and an isolate of C. beticola from Montana (GenBank Accession No. AF443281); the sequences were 94.6% similar, an appreciable difference. For pathogenicity tests, cultures were grown on carrot leaf decoction agar. Aqueous suspensions of 104 spores per ml from cultures were sprayed on 6-week-old L. draba plants. Plants were covered with plastic bags and placed on the greenhouse bench at 20 to 25°C for 96 h. Koch's postulates were completed by reisolating the fungus from the circular leaf spots that appeared within 10 days, usually on lower leaves. Spores of C. bizzozeriana were also sprayed on seedlings of sugar beet, collard, mustard, radish, cabbage, and kale under conditions identical to those above. No symptoms occurred. After the discovery of the disease in 1997, plants of L. draba in eastern Montana, Wyoming, and Utah were surveyed from 1998 to 2003 for similar symptoms and signs, but none were found. This, to our knowledge, is the first report of C. bizzozeriana in the United States. The initial report of the fungus in North America was from Manitoba in 1938 (1). It has recently been reported as occurring on L. draba in Tunisia (4) and Russia (3) and is reported as common in Europe (2). A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI No. 878750A). References: (1) G. R. Bisby. The Fungi of Manitoba and Saskatchewan. Natl. Res. Council of Canada, Ottawa, 1938. (2) C. Chupp. A Monograph of the Fungus Genus Cercospora. C. Chupp, Ithaca, NY, 1953. (3) Z. Mukhina et al. Plant Dis. 92:316, 2008. (4) T. Souissi et al. Plant Dis. 89:206, 2005.

Leafy spurge (Euphorbia esula-virgata), a native of Eurasia, is a serious invasive weed of western grasslands of North America. It is very difficult and cost-prohibitive to control with herbicides; control by insect biological control agents and cultural practices are minimally effective in suppressing vegetative growth and seed production. Current biological control of leafy spurge with pathogens is primarily with mycoherbicides, which require specific environmental conditions and repeated applications to be effective. Alternative biological control approaches using selected microorganisms to attack roots and adventitious shoots may effectively decrease vigor of leafy spurge without environmental manipulations to ensure control efficacy. Our objectives were to survey leafy spurge accessions and their native soils for associated microorganisms, and to assess these microorganisms for potential biological control. A proposed screening protocol, consisting of primary and secondary assays, effectively detected bacteria and fungi with high phytotoxicity toward leafy spurge. Primary lettuce seedling bioassays indicated that 62 % of rhizosphere and 54 % of endorhizal bacteria significantly (P = 0.05) inhibited root growth, causing necrotic lesions. Over 60 % of fungal isolates bioassayed on rice agar significantly inhibited root growth of lettuce seedlings. The most effective microbial isolates, based on primary bioassays, were screened directly on leafy spurge cuttings in secondary assays. Only intact cells of bacteria were detrimental to leafy spurge, indicating that host-bacterial contact was required for pathogenicity. Culture filtrates of 40 % of the test fungi caused complete chlorosis and leaf wilting. The most effective fungi originated from leafy spurge adventitious shoots. Results of the survey suggest that leafy spurge rhizospheres and adventitious shoots are potential sources of biological control microorganisms, which should be considered for inclusion in comprehensive management programs for leafy spurge.

Summary The fluorescent antibody technique and the immunodiffusion test were used for the serological detection of type C virus found in one bovine and two sheep established cell lines. The electron microscopic demonstration of type C virus particles supported by uridine labelling suggested that a virus specific antigen underlay the FA reaction and the reaction of identity in the IDT. Preliminary FA tests employing various antisera against mammalian oncornaviruses may indicate that bovine leukemia virus represents antigenically a unique group. Zusammenfassung Untersuchungen uber die Rinderleukose I. Gewinnung von C-Typ virusproduzierenden Zell-Linien Leukozyten einer Leukose-kranken Kuh und Lymphknoten-explantate von zwei Leukose-infizierten Schafen dienten zur Gewinnung permanenter Zell-Linien. In diesen Zellkulturen konnten C-Typ Virus und Virusantigen elektronenoptisch, fluoreszenzserologisch, mittels Immunodiffusion und durch Markierung mit 3H-Uridin nachgewiesen werden. Vergleichende fluoreszenzserologische Untersuchungen mit Seren gegen eine Reihe von Oncornaviren anderer Saugetiere deuten darauf hin, das das Virus der Rinderleukose eine antigen selbstandige Untergruppe der Oncornaviren darstellt. Resume Recherches sur la leucose bovine I. Obtention des lignees cellulaires produisant du virus type C Des leucocytes d'une vache atteinte de leucose et des explantats de ganglions de deux moustons infectes de leucose ont servi a obtenir les lignees cellulaires permanentes. On a pu mettre en evidence dans ces cultures cellulaires un virus du type C et un antigene viral au microscope electronique, par l'immunofluorescence, au moyen de l'immunodiffusion et par marquage avec 3H-uridine. Des recherches comparees en fluorescence avec des antiserums d'une serie d'oncornavirus d'autres mammiferes ont montre que le virus de la leucose bovine represente un sous-groupe antigeniquement unique. Resumen Estudios sobre la leucosis bovina I. Obtencion de lineas celulares que producen el virus tipo C Los leucocitos de una vaca leucotica y las explantaciones ganglionares de dos ovejas infectadas con leucosis se utilizaron para obtener lineas celulares permanentes. En estos cultivos celulares se identificaron particulas virales del tipo C con la ayuda del microscopio electronico y del marcaje con uridina-3H. Se detectaron tambien antigenos virales con las tecnicas de immunofluorescencia e imunodifusion. Los estudios fluorescenteserologicos comparativos realizados con sueros contra una serie de virus oncorna de otros mamiferos parecen indicar que el virus de la leucosis pertenece a un subgrupo antigenicamente independiente de los virus oncorna.

In order to select compatible human leucocytes antigens (HLA) donors for bone marrow graft, all the members of 76 families were typed by serology for HLA class I (A and B locus) and class II (DR, DQ locus) by polymerase chain-reaction-sequence-specific primes (PCR-SSP). The HLA typing interpretation revealed the existence of crossing-over in major histocompatibility (CMH) regions for two families, AB and AT, with aplastic bone marrow. The study of crossing-over site has needed the genotyping of seven short tandem repeat (STR) markers located on the short arm of chromosome 6 (D6S291, D6S273, TNFa, C1.2.C, C3.2.11, D6S265, D6S276), using ABI Prism 310 sequencer. HLA and STR Haplotypic analysis enabled us to confirm the crossing-over between locus B and DR in AB family and between locus A and B in AT family. Based in this study, we recommend to be careful in the interpretation of the results of HLA typing between donors and recipients of bone marrow. Complementary investigations should be accomplished for studying genetic abnormalities, which would be involved in this pathology.

The occurrence of an isolated ovarian or pelvic relapse of acute lymphoblastic leukemia (ALL) in complete remission after chemotherapy has rarely been described. We report the case of a 12-year-old girl, treated for ALL, who developed an isolated left ovarian and fallopian tube localization without medullary or blood relapse 4 years after the end of the initial treatment. She presented with an isolated pelvic mass. The diagnosis was established by a CT-guided biopsy. The treatment consisted of a second course of chemotherapy and complementary surgery; a second complete remission was obtained.

Resume Introduction Les immunosuppresseurs sont habituellement indiques dans le traitement de premiere ligne des formes non severes de l’aplasie medullaire acquise (AMA). Le but de ce travail etait d’evaluer l’efficacite d’un protocole prospectif associant la ciclosporine aux androgenes dans le traitement des AMA non severes de l’adulte. Methodes Vingt-neuf cas diagnostiques et suivis a Sfax ont ete inclus sur une periode de dix ans (1991–2000). Mis a part le traitement symptomatique, tous nos patients ont recu un traitement specifique comportant une bitherapie a base de ciclosporine a la dose de 5 mg/kg par jour et des androgenes a la dose de 0,5 mg/kg par jour. Resultats Les taux de reponses a trois mois, a six mois et a 12 mois sont respectivement de 48, 75 et 87 %. Les taux de survies sont de 52 % a un an et de 37 % a cinq et dix ans. Les principales toxicites etaient hepatiques et renales ainsi que l’hypertension arterielle, qui ont ete observees chez respectivement 53, 16 et 15 % des patients avec une reversibilite respective dans 65, 87 et 100 % des cas. Conclusion Les taux de reponse et de survie dans notre serie sont assez satisfaisants comparativement a ceux obtenus par les immunosuppresseurs (serum antilymphocytaire et ciclosporine) dans la litterature. L’association des androgenes a permis de potentialiser l’immunosuppression induite par la ciclosporine mais au prix d’effets secondaires plus importants.

Resume A propos d'une observation d'aplasie medullaire avec syndrome malformatif nous discutons les criteres diagnostiques et les cadres nosologiques des 2 principales aplasies medullaires hereditaires s'accompagnant d'un syndrome malformatif : la maladie de Fanconi et la dyskeratose congenitale. Observation Une jeune fille, âgee de 19 ans, issue d'un mariage consanguin au troisieme degre, a ete hospitalisee pour exploration d'un syndrome anemique et hemorragique. L'examen clinique a montre un syndrome malformatif associant une microphtalmie, des taches cafe au lait, une hyperpigmentation generalisee et une dystrophie ungueale des doigts et des orteils sans leucoplasie des muqueuses. Un retard staturoponderal a ete note. L'hemogramme a montre une pancytopenie et la biopsie medullaire une moelle hematopoietique desertique en faveur d'une aplasie medullaire. Le caryotype realise sur des lymphocytes sanguins apres sensibilisation par la mitomycine C, a objective un syndrome d'instabilite chromosomique franc. Le diagnostic d'aplasie medullaire hereditaire a ete retenu et la patiente a ete mise sous norethandrolone. La patiente est decedee3 mois plus tard par choc septique. Discussion Devant un tableau d'aplasie medullaire associe a un syndrome malformatif, le diagnostic d'aplasie medullaire hereditaire est le plus probable. La maladie de Fanconi est la plus frequente, evoquee devant une dysmorphie de la face caracteristique avec microphtalmie, des taches cafe au lait, un retard staturoponderal, des anomalies de la colonne du pouce. La dystrophie ungueale, la leucoplasie des muqueuses sont presque pathognomoniques du diagnostic de dyskeratose congenitale. Si le syndrome malformatif est ambigu, l'etude cytogenetique ne permet pas de lever le doute diagnostique dans toutes les situations, comme dans notre observation.

Factor XIII deficiency is a rare autosomal recessive congenital disorder of haemostasis characterised by a plasmatic factor XIII level less than 1% in homozygote and bleeding as of the youth. We report a study about ten patients with congenital factor XIII deficiency from seven south-Tunisian families, there are seven females. Umbilical bleeding was common and only two patients had intracranial bleeding. The standard screening tests are normal. Factor XIII activity was less than 1% in all patients. A sub-unit A deficit was detected for the ten patients. Out hemorrhagic context, five patients receive regular prophylactic transfusion with fresh frozen plasma.

Resume La lymphadenite subaigue de Kikuchi-Fujimoto ou maladie de Kikuchi est une entite anatomoclinique d'evolution benigne de l'adulte jeune. Nous rapportons l'observation d'une jeune fille âgee de 20 ans consultee pour syndrome polyganglionnaire associe a une fievre. L'examen a revele des adenopathies disseminees faisant jusqu'a 6 cm de diametre. La vitesse de sedimentation etait acceleree a 40 mm a la premiere heure, les lacticodeshydrogenases etaient elevees a 593 UI/l. Les serologies de la rubeole, de l'hepatite B et C, du VIH et de la toxoplasmose etaient negatives ainsi que le bilan tuberculeux. La biopsie d'un ganglion cervical a conclu a une adenite necrosante non suppuree contenant des debris nucleaires avec une importante reaction histiocytaire, aspect en faveur d'une maladie de Kikuchi. L'evolution apres trois mois etait marquee par la regression des adenopathies sans aucun traitement. Il s'agit d'une nouvelle observation de maladie de Kikuchi caracterisee par un tableau clinicobiologique generalise, faisant evoquer fortement une hemopathie maligne en particulier lymphomateuse.

The input costs of pesticides to control biotic constraints are often prohibitive to the subsistence farmers of Africa and seed based solutions to biotic stresses are more appropriate. Plant breeding has been highly successful in dealing with many pest problems in Africa, especially diseases, but is limited to the genes available within the crop genome. Years of breeding and studying cultural practices have not always been successful in alleviating many problems that biotechnology may be able to solve. We pinpoint the major intractable regional problems as: (1) weeds: parasitic weeds (Striga and Orobanche spp.) throughout Africa; grass weeds of wheat (Bromus and Lolium) intractable to herbicides in North Africa; (2) insect and diseases: stem borers and post-harvest grain weevils in sub-Saharan Africa; Bemesia tabaci (white fly) as the vector of the tomato leaf curl virus complex on vegetable crops in North Africa; and (3) the mycotoxins: fumonisins and aflatoxins in stored grains. Abiotic stresses may exacerbate many of these problems, and biotechnological alleviations of abiotic stress could partially allay some predicaments. Some of these constraints are already under study using biotechnological procedures, but others may require longer-term research and development to alleviate the problems. Despite the huge impacts of post-harvest weevils and of mycotoxins in grains, these issues had not been given high priority in national biotechnological programs, possibly due to a lack of knowledge of their immensity. The need for public sector involvement is accentuated for cases where immediate profits are not perceived (e.g. lowering mycotoxin levels in farmer utilized grain, which does not increase yield) but where the public weal will gain, and will be invaluable, especially where the private sector supplies genes already isolated.

Resume Objectifs . — Degager les aspects epidemiologiques, cliniques, biologiques et evolutifs de la maladie de Fanconi dans le sud tunisien. Patients et methodes . — Nous avons collige 43 cas sur une periode de 12 ans. Pour chaque malade ont ete realises un examen clinique, un hemogramme, un myelogramme, une biopsie medullaire, une electrophorese de l’hemoglobine et un caryotype, une radiographie du squelette, une echographie abdominale et une UIV. Tous les malades au stade de pancytopenie etaient traites par des androgenes. Aucun malade n’a beneficie d’une allogreffe de moelle osseuse. Resultats . — Vingt-quatre filles et 19 garcons (âge moyen de 10 ans et 9 mois). Le caractere familial a ete retrouve dans 53% des cas. Le motif de consultation le plus frequent etait un syndrome anemique (69%). Le diagnostic a ete pose lors d’une enquete familiale dans dix cas (24%). Le syndrome malformatif etait present dans tous les cas (troubles de pigmentation: 86%; retard de maturation osseuse: 83%; dysmorphie faciale: 76%; hypotrophie staturale: 65%; anomalies osseuses: 53%; malformations renales: 44%). L’anemie a ete retrouvee dans 88% des cas. La thrombopenie et la neutropenie etaient constantes. La moelle etait, dans tous les cas, hypoplasique ou aplasique a la biopsie medullaire. Des cassures chromosomiques spontanees ont ete retrouvees dans 79% des cas etudies. Le taux d’hemoglobine fœtale etait eleve dans 80% des cas etudies avec un taux moyen de 20,5%. Le taux de survie actuarielle a 5 ans etait de 48%, mais les longues survies etaient rares (huit patients sur 43). Conclusions : — Cette maladie parait frequente dans le Sud tunisien. Un caryotype normal selon les techniques classiques, retrouve chez cinq de nos patients, ne peut pas ecarter le diagnostic, d’ou l’interet des agents sensibilisants. Par ailleurs, un taux anormal d’hemoglobine foetale a permis, dans certains cas, d’evoquer le diagnostic. l’androgenotherapie a permis d’accroitre la survie a plus de 5 ans chez huit patients. Cependant, le seul espoir de guerison reste la greffe de moelle osseuse.

Screening large collections of microorganisms for potential biological control activity on economically important weeds is often diY cult. Seeds required as indicator species are often not available in large supply and are highly variable in viability and germination, resulting in uneven seedling development. A bioassay system for rapidly assessing the phytotoxic eVects of rhizobacteria was developed based on leafy spurge ( Euphorbia esula L. ) callus tissue culture in multiple-well plates. Callus pieces (0.5 g) were placed in 24-well plates containing Gamborg's B5 medium, inoculated with rhizobacterial suspension and incubated at 27oC for 48 h. By rating inoculated callus for cellular damage, about 30% of rhizobacteria isolated from weedy Euphorbia spp. collected in Europe and North America were identified as being highly phytotoxic. Symptoms of phytotoxicity included growth reduction, discoloration and extensive cellular leakage. A high proportion of isolates found to be phytotoxic in tissue culture bio...

The occurrence of an isolated ovarian or pelvic relapse of acute lymphoblastic leukemia (ALL) in complete remission after chemotherapy has rarely been described. We report the case of a 12-year-old girl, treated for ALL, who developed an isolated left ovarian and fallopian tube localization without medullary or blood relapse 4 years after the end of the initial treatment. She presented with an isolated pelvic mass. The diagnosis was established by a CT-guided biopsy. The treatment consisted of a second course of chemotherapy and complementary surgery; a second complete remission was obtained.

Immunosuppressive drugs are usually used in the treatment of acquired aplastic anemia (AAA). The aim of this study was to evaluate the efficiency of a prospective therapeutic protocol using cyclosporine and androgens in the treatment of adult patients with nonsevere AAA.Twenty-nine patients diagnosed and treated at the University Hospital of Sfax (Tunisia), during a 10-year period (1991-2000) were included. In addition to symptomatic treatment (transfusion, antibiotics), all the patients received a specific treatment including two drugs: cyclosporine 5mg/kg per day and androgens 0,5mg/kg per day.The response rate at three, six and 12 months were 48, 75, and 87%, respectively. Survival rate was 52% at one year, and 37% at five and 10 years. The main toxicities were hepatic, renal, and hypertension, observed in 53, 16 and 15%, respectively. These toxicities were reversible in 65, 87 and 100% of the cases, respectively.The response and survival rates in our series are quite satisfactory when compared to those obtained with other immunosuppressive drugs (cyclosporine and antilymphocyte serum) in the literature. The addition of androgens in our patients seemed to potentiate the immunosuppression induced by ciclosporin, but secondary toxic effects were more common.

In order to select compatible human leucocytes antigens (HLA) donors for bone marrow graft, all the members of 76 families were typed by serology for HLA class I (A and B locus) and class II (DR, DQ locus) by polymerase chain-reaction-sequence-specific primes (PCR-SSP). The HLA typing interpretation revealed the existence of crossing-over in major histocompatibility (CMH) regions for two families, AB and AT, with aplastic bone marrow. The study of crossing-over site has needed the genotyping of seven short tandem repeat (STR) markers located on the short arm of chromosome 6 (D6S291, D6S273, TNFa, C1.2.C, C3.2.11, D6S265, D6S276), using ABI Prism 310 sequencer. HLA and STR Haplotypic analysis enabled us to confirm the crossing-over between locus B and DR in AB family and between locus A and B in AT family. Based in this study, we recommend to be careful in the interpretation of the results of HLA typing between donors and recipients of bone marrow. Complementary investigations should be accomplished for studying genetic abnormalities, which would be involved in this pathology.

Based on a case report of aplastic anemia associated with malformation, we discuss the diagnostic criteria and the nosologic problem between the 2 principal aplastic anemia accompanied with malformation: Fanconi disease and dyskeratosis congenita.A 19-year-old girl, issued from a third degree consanguineous marriage, was admitted because of anemic and hemorrhagic syndrome. Physical examination showed several malformations: microphtalmia, brownish spots, generalized hyperpigmentation and ungueal dystrophy without mucosal leucoplasia. Statural and ponderal retardation were noted. On the hemogram there was a pancytopenia and on biopsy, the bone marrow was desertic. The caryotype performed on peripheral blood lymphocytes after sensibilisation with mitomycin C revealed chromosomal instability aspects. Based on these clinical and biological features, the diagnosis of hereditary aplastic anaemia was retained. The patient was given norethandrolone. She died 3 months later by septic shock.Coexistence of aplastic anemia with a malformative syndrome suggests most probably an hereditary form of aplastic anemia. Fanconi anemia is the most frequent. It associates characteristic anomalies of the face, with microphtalmia, brownish spots, statural and ponderal retardation, and thumb anomalies. Ungueal dystrophy, mucosal leucoplasia are almost pathognomonic of congenital dyskeratosis. When the malformative syndrome is not characteristic, the cytogenetic study may also fail to make the differential diagnosis, as was the situation in our case.

Factor XIII deficiency is a rare autosomal recessive congenital disorder of haemostasis characterised by a plasmatic factor XIII level less than 1% in homozygote and bleeding as of the youth. We report a study about ten patients with congenital factor XIII deficiency from seven south-Tunisian families, there are seven females. Umbilical bleeding was common and only two patients had intracranial bleeding. The standard screening tests are normal. Factor XIII activity was less than 1% in all patients. A sub-unit A deficit was detected for the ten patients. Out hemorrhagic context, five patients receive regular prophylactic transfusion with fresh frozen plasma.

Subacute necrotizing lymphadenitis or Kikuchi–Fujimoto disease is a benign pathological entity diagnosed chiefly in young adults. We report a case in a 20-year-old woman who presented with swollen lymph nodes and a fever. Disseminated lymphadenopathy with nodes measuring up to 6 cm in diameter was found upon physical examination. The erythrocyte sedimentation rate was elevated to 40 mm/h and the lactic dehydrogenase level to 593 IU/l. Findings were negative from serological tests for rubella, hepatitis B, hepatitis C, HIV, and toxoplasmosis, as well as from tests for tuberculosis. A biopsy of a cervical lymph node showed nonsuppurative necrosis, karyorrhexis, and a marked histiocytic reaction consistent with Kikuchi–Fujimoto disease. No treatment was given, and a full recovery was achieved within 3 months. In this patient, the disseminated lymphadenopathy and constitutional symptoms strongly suggested a hematological malignancy and more specifically a lymphoproliferative disease.

We report the case of a 8-year-old girl diagnosed with myelodysplastic syndrome. This case was morphologically characterized by the presence of bundle of Auer rods in the neutrophils. The evolution of the disease was marked by a quick transformation in a acute myeloid leukaemia with t(8;21) refractory to treatment. We reviewed the literature for clinical, biological and therapeutic features of this rare childhood hemopathy.

Pleural effusion caused by plasma cell involvement in multiple myeloma has been reported unfrequently, and has been described at a frequency below 1% of multiple myeloma. In this study, we report an observation with pleural effusion as first symptom of multiple myeloma. The analysis of the pleural liquid showed plasma cells with a monoclonal IgG Kappa immunoglobulin. In addition, there was a bone marrow infiltration by plasma cells, a serum monoclonal immuno-globulin of the same type and osteolytic lesions. Our patient has received one course of chemotherapy with: vincristine, melphalan, cyclophosphamide and prednisone. The patient did not respond to the therapy and died one month later. Pleural effusion seems to be an expression of aggressive myeloma. Survival exceeds rarely 4 months.

We report a retrospective study of 42 cases of lymph node tuberculosis. We noted symptoms of tuberculosis impregnation in 92%, cervical localization in 71%, positive tuberculin intra-dermo-reaction in 77%, and accelerated erythrocyte sedimentation rate in 73% of the cases. Koch bacillus was detected in expectoration, urine or gastric liquid at the rate of 11% of the cases. Lymph node function was suggestive in 4 out of 12 patients, showing giant cells with or without caseum. Lymph node biopsy, performed in 32 patients, was contributive in 94% of them. Another tuberculous localization was found in 14 cases mainly pulmonary (8 cases).

Hemoglobin abnormalities constitute a public health problem in many countries in the world. In Tunisia, these disorders were thought to affect only the North-western population. However, the existence of hemoglobinosis concentration in Kebily in south Tunisia has been suggested by previous work. In order to estimate their frequencies, we performed a screening of hemoglobin abnormalities in the North-Kebili region, to establish a prevention program of the homozygous forms.This screening concerned all 1st and 2nd grade primary school pupils in North Kebily. After a questionnaire, a blood sample was drawn from every child. Hemogram, sickling test, and hemoglobin electrophoresis at alkaline pH were performed for all children. Hemoglobin electrophoresis at acid pH and a specific hemoglobin A2 titration were performed for some children.The study concerned 1,400 children, aged between 5 and 12 years, the mean age was 7 years and 7 months +/- 10 months. Consanguinity rate and coefficient were respectively 44% and 2249 x 10(5). Endogamy was very high. The global rate of hemoglobin abnormalities was 9.4%. Drepanocytosis with a rate of 4.9% was the most frequent, followed by beta thalassemia (3.1%) and C hemoglobinosis (1.6%). These abnormalities were unequally distributed; very frequent in some localities, they were quite absent in others.This study revealed a hemoglobinosis concentration in Tunisia, which can be classified second after that of Beja in North-western Tunisia. The heterogeneous distribution of the hemoglobin abnormalities in North-Kebili region and the high consanguinity and endogamy rates constitute factors that promote homozygous and double heterozygous forms to arise and justify the elaboration of a preventive strategy.

To report the epidemiologic, clinical, biological features and course of Fanconi's anemia in southern Tunisia.During a period of 12 years we observed 43 cases. For each patient, careful clinical, biological (hemogram, myelogram, bone marrow biopsy, hemoglobin electrophoresis, karyotype) and radiological (skeleton X-rays, abdominal echography and intravenous urography) examinations were performed. All the patients who were at a pancytopenia stage were given androgens. None had a bone marrow allograft.There were 24 girls and 19 boys. The mean age at diagnosis was 10 years and 9 months. The familial character was present in 53% of the cases. The most frequent initial complaint was anemic syndrome (69%). In ten cases (24%), the diagnosis has been established during a familial investigation. Malformations were present in all cases (abnormal pigmentation: 86%; skeletal maturation retardation: 83%; facial dysmorphy: 76%; statural hypotrophy: 65%; bone abnormalities: 53%; renal malformations: 44%). Anemia was present in 88% of the cases, thrombocytopenia and neutropenia in all cases. Bone marrow was hypoplastic or aplastic in all cases on biopsies. Spontaneous chromosomal breaks were found in 79% of the studied cases. Fetal hemoglobin was increased in 80% of the studied cases with a mean level of 20.5%. Actuarial survival rate at 5 years was 48%, but long survival durations were rare (eight out of 43 patients).This disease, rare in the world, seems to be frequent in southern Tunisia. A normal karyotype (with classical techniques), found in five patients, could not discard the diagnosis; for this reason, the use of sensitizing agents should improve the sensitivity of the test. Besides, an increased level of fetal hemoglobin enabled us to suggest the diagnosis in some cases. Androgenotherapy increased the survival duration to more than 5 years in eight patients. However, bone marrow allograft remains the only possibility of cure.

Sickle cell anemia is a monogenic hereditary disease characterized by a mutation in the beta globin gene. Five major haplotypes associated with the beta S mutation have been defined: Benin, Bantu, Senegalian, Camerounian, and Arabo-Indian. Previous studies in northern Tunisia showed that sickle cell anemia was of Benin origin in this region. Patients from the south of Tunisia, mainly from the Kebili region, were not previously concerned. In this study, we have determined the beta S haplotype and evaluated phenotypical expression of the disease in 14 patients from this latter region. The use of four restriction endonucleases having polymorphic sites in the beta globin gene showed that all patients had the Benin haplotype, confirming the Benin origin of sickle cell anemia in Tunisia. This haplotype is associated with an heterogeneous expression of fetal hemoglobin (HbF) with extremes varying from 2.4 to 16.3% and a mean expression rate of 8.16%, which is in accordance with literature data. In spite of the haplotype homogeneity in our patients, clinical heterogeneity was noted. A unique case of alpha-thalassemia could not explain this heterogeneity. In contrast, we found a certain correlation between fetal hemoglobin expression and clinical severity.

Between January 1994 and June 1995, 19 cases of nasopharyngeal carcinoma, classified N2-N3 (AJC/UICC 1987) were treated by neoadjuvant chemotherapy including cisplatin 100 mg/m2 on day 1 and epirubicin 80 mg/m2 on day 1. Following course started on day 21. A clinical and scannographic evaluation was made after 3 courses of chemotherapy. Fifty-eight percent of the patients were N3. Seventy-four percent were T3-T4. Tolerance to chemotherapy was good in 100% of the cases. A functional improvement was obtained in 14 among 16 patients who were initially symptomatic. For lymph nodes, an objective response (OR) was observed in all patients. The response was complete (CR) in 53%. A regression of primary tumor was obtained in 68% of the patients, but it was complete in 16% only. Sixty-nine percent of the patients has have a tumoral OR have a lymph node CR. Ninety percent of the patients with a complete lymph node response have a tumoral regression more than 50%. A correlation seems likely between the importance of the tumoral and the lymph node responses.

This is a retrospective study on the use of cytarabine at low doses in acute myeloid leukemias in 41 patients. Four groups of AML are included: group A: 19 cases of de novo AML in elderly patients; group B: ten cases of AML in relapse; group C: five cases of AML refractory to previous treatment and group D: seven cases of secondary AML. Cytarabine was given subcutaneously at the dose of 10 mg/m2 of body surface, for 21 days per month for the first course; then 15 days per month for the following courses. The response rates were 42, 20, 0, and 43% respectively for the A, B, C, and D groups. A complete remission was attained in only 15% (six patients). Extra haematological tolerance was excellent. Infection complications were noted in 66%, whereas a severe neutropenia was observed in 34% of patients. Hemorrhagic complications were more rare (20% of patients). The mean duration of complete remission was 10 months. The median survival was 10.5 months (2 to 31 months) for the responder patients, and 2.4 months (1 to 7 months) for the non-responders. Cytarabine at low doses seems to be a good indication for first intention treatment of AML in elderly patients. It does not give a bone marrow aplasia, the infection and hemorrhagic episodes are less numerous than with conventional dose chemotherapy, the life quality is improved, and treatment at home is often possible.

Aplastic anaemia is a potentially fatal haematopoietic disorder whose aetiology is not yet clarified. In our preliminary study we have introduced cyclosporin in the aplastic anaemia treatment to evaluate its effect on the disease evolution. Ten aplastic anaemia patients, mean age 33.33 +/- 20.01 years, were treated with cyclosporine (9 +/- 2.35 mg/kg/d), prednisolone (0.5 mg/kg/d) and androgens (1 mg/kg/d). The prednisolone was always combined with cyclosporine. The androgens were administered concomitantly with the cyclosporine or alternately. Seven patients responded to the treatment after a median remission delay of 6 weeks (2-12 weeks). They became independent of blood requirements at a median of 36 weeks (8-108 weeks); the three other patients died during the first trimester without showing any improvement. Among the seven responders, two relapsed early and transiently. The rate of actuarial survival was 70 per cent. The median duration of survival was 10.5 months. The side effects observed included one case of malignant lymphoma, six cases of liver toxicity and five cases of kidney toxicity. This toxicity was reversible after dose adjustment of the cyclosporine. In our study, the introduction of cyclosporin in the aplastic anaemia treatment resulted in improved therapeutic response. Androgens should be used to maintain the haematologic response. This therapeutic protocol associated with drug monitoring seems promising and the side effects should not limit its use because of the severity of the underlying disease.

The authors report a rare case of regional portal hypertension secondary to splenic vein obstruction by left adrenal carcinoma metastasis. They review the principal literature data concerning regional portal hypertension and adrenal cortical carcinoma.

Silybum eburneum Coss. & Durieu. (ivory thistle) and S. marianum (L.) Gaertn. (milk thistle) are dominant, invasive weeds in northern Tunisia (1). S. marianum is also invasive in the United States and targeted for biological control. The smut fungus Microbotryum silybum Vánky & Berner is a naturally occurring pathogen of S. marianum in Greece (2) but not in Tunisia or the United States. To assess the safety of the fungus for biological control in the United States, plants related to S. marianum were evaluated for susceptibility to M. silybum in the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. Because of the close genetic relationship of S. eburneum to S. marianum, both were tested for susceptibility under greenhouse conditions at the FDWSRU. All inoculations were done by placing 5 mg of teliospores of M. silybum in the central whorl of rosettes with three to five true leaves. Individual plants in soil-filled pots were placed in a controlled chamber at 16°C with 10 h of light daily. Photon flux density in the chamber was 34 μmol·m-2·s-1 supplied by three 1.8-m long 115W fluorescent tubes and three 52W incandescent bulbs. The central whorl was misted with distilled water twice daily for 2 weeks and the temperature was then lowered to 8°C for 6 weeks. The plants were transferred to a greenhouse bench at 22 to 25°C with 14 h of light daily. Photon flux density on the bench was 620 μmol·m-2·s-1 provided by two 500W sodium vapor lamps, one 1,000W metal halide lamp, and incidental sunlight. After approximately 7 weeks, plants of each species had fully developed capitula that flowered normally, produced no flowers, or formed abnormal flowers. Abnormal capitula contained powdery masses of teliospores in the ovaries of the florets. In contrast to systemic infections that were observed in the field (2), different branches of bolted plants bore both diseased and normal capitula. In turn, diseased capitula of both species were either completely diseased (all florets filled with teliospores) or partially diseased. Four of ten S. marianum plants and six of nine S. eburneum plants were diseased. Pathogenicity tests were repeated four times with similar results. In Greece, field inoculation of S. marianum with 5 mg of teliospores produced an average of 89% diseased plants with an average of 250 g of teliospores produced per plant. A similar level of disease is possible for S. eburneum under field conditions. Teliospores from smutted ovaries of both plant species conformed to the description for M. silybum (2). Both species are annual plants that reproduce solely by seeds. Since M. silybum prevents seed production, this fungus has great potential as a biological control agent in the United States and Tunisia. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI 863477). Nucleotide sequences for the internal transcribed spacer region are available in GenBank (Accession No. AY285774). To our knowledge, this is the first report of M. silybum parasitizing S. eburneum. References: (1) G. Pottier-AlaPetite. Flore de la Tunisie: Angiospermes-Dicotylédones, Gamopétales, Tunis, 1981. (2) K. Vánky and D. Berner. Mycotaxon 85:307, 2003.

Lepidium draba (L.) subsp. draba (synonym = Cardaria draba (L.) Desv.), commonly known as white-top or hoary-cress (1), family Brassicaceae, is a common weed and emerging problem in wheat in Tunisia. It is also a problematic invasive weed in the northwestern United States and a target of biological control efforts. During the summer of 2002, dying L. draba plants were found around Tunis, Tunisia. Plants had grayish white leaf spots on most of the leaves. In some cases, the leaf spots dropped out of the leaves producing “shot-holes”. In most cases, the leaf spots coalesced, and the leaves wilted and died. Diseased leaves were collected, air-dried, and sent to the quarantine facility of the Foreign Disease-Weed Science Research Unit (FDWSRU), USDA/ARS, Fort Detrick, MD. The air-dried leaves were observed microscopically, and numerous conidiophores and conidia were observed on both sides of the leaves within and around the lesions. The fungus isolated (DB03-009) conformed to the description of Cercospora bizzozeriana Saccardo & Berlese (2). Conidiophores were unbranched, pale olive-brown, 1 to 5 geniculate, and uniform in color and width. Conidia were hyaline, straight to slightly curved, multiseptate, and 57 to 171 × 3.8 to 6.7 µm (average 103 to 4.6 µm). Stems and leaves of 12 rosettes (10 to 15 cm in diameter) of 6-week-old L. draba plants were spray inoculated with an aqueous suspension of conidia (1 × 105/ml) harvested from 6- to 8-day-old cultures grown on carrot leaf decoction agar. Six of the plants and two noninoculated plants were placed in a dew chamber at 22°C in darkness and continuous dew. The other half of the plants and two noninoculated plants were placed on a greenhouse bench at approximately 25°C and covered with clear polyethylene bags. After 72 h, plants from the dew chamber were moved to a greenhouse bench, and the bagged plants were uncovered. All plants were watered twice daily. After 9 days, symptoms were observed on the plants that had been bagged but not on the plants from the dew chamber. Symptoms were identical to those observed in the field in Tunisia and included “shot holes”. No symptoms were observed on noninoculated plants. C. bizzozeriana was reisolated from the leaves of all symptomatic plants. Completion of Koch's postulates was repeated with an additional five plants. This isolate of C. bizzozeriana is a destructive pathogen on L. draba subsp. draba, and severe disease can be produced by inoculation of foliage with an aqueous suspension of conidia. This isolate is a good candidate for mycoherbicide development in Tunisia where the weed and pathogen are indigenous. However, some commercially grown Brassica species were found susceptible to this isolate, which will preclude its use as a classical biological control agent in the United States. To our knowledge, this is the first report of C. bizzozeriana on L. draba subsp. draba in Tunisia. A voucher specimen has been deposited at the U.S. National Fungus Collections (BPI 843753). Live cultures are being maintained at FDWSRU and the Institut National Agronomique de Tunisie, Tunis, Tunisia. References: (1) I. A. Al-Shehbaz and K. Mummenhoff. Novon 12:5, 2002. (2) C. Chupp. A Monograph of the Fungus Genus Cercospora. C. Chupp, Ithaca, New York, 1953.

Pesticides are used extensively in ornamental production. Studies of repeated pesticide applications indicate that microbial changes occur in the rhizosphere of the plant. In addition to controlling the target pest, often a population shift of bacteria may occur. This has been previously shown in research associated with leatherleaf fern [Rumohra adiantiformis (Forst.) Ching] and the fungicide benomyl. Rhizobacteria (root-associated bacteria) of anthurium (Anthurium andraeanum) were investigated with respect to total populations and isolates that are potentially phytotoxic. The anthurium sample roots were taken from commercial Jamaican production sites. The sites had either a benomyl or non-benomyl history. Rhizobacterial populations were estimated by dilution plating and subcultures were taken for a phytotoxicity bioassay. Micrographs of samples were prepared to examine treatment effects on the morphology of roots. Rhizobacteria populations were frequently at 106 colony forming units per gram fresh weight. Consistently, greater than 50% of the isolates from each treatment were potentially phytotoxic. However, in the benomyl history samples, there was a greater diversity of phytotoxic isolates.

The [ 125 I ] Clq-binding activity (Clq BA) and the antibody titre against Epstein-Barr (EB) viral capsid antigen (VCA), EB nuclear antigen (EBNA) and early antigen (EA) were determined in sera from 28 nasopharyngeal carcinoma patients and 66 matched controls from Tunisia. The geometric mean titres (GMT) of anti-VCA IgG and anti-EBNA, as well as the prevalence of anti-EA, antibodies were significantly higher among the patients than in the control group. The increased Clq BA of NPC sera, as compared to that of control sera, is likely to reflect the presence of soluble immune complexes, to which the contribution of the various autoantibodies found in this disease seems to be negligible. In individual sera from these patients, the Clq BA correlated positively with the titre of IgG ( P = 0.026), but not with that of IgA, anti-VCA antibodies, indicating that the age-dependency of these different parameters was not per se a sufficient condition for a positive correlation to appear. Since the correlation between Clq BA and anti-VCA titre was restricted to the class of Ig which binds Clq, an alternative explanation is proposed in which specific anti-EBV antibodies complexed to virion structural antigens account for part of the elevated Clq BA.

Various facts are now known about the relative lymphoma resistance of a group of tetraparental AKR reversible CBA/H-T6 chimaeras derived by early embryo aggregation. Firstly, their tumour resistance is not due to the lack of the lymphomaprone AKR cells. Secondly, results showing titres of MuLV-gs antigen comparable with, and occasionally in excess of, those in the AKR suggest that the tumour resistance of the chimaeras is unlikely to be due to a lack of oncogenic leukaemia virus. However, in marked contrast to the AKR, antibody-viral antigen renal complexes in the chimaeras were minimal. Lack of viral antigens could not explain the relative lack of renal complexes. Absence of the corresponding anti-viral antibody is the most likely explanation and this has to be attributed to the CBA component of the tetraparental AKR reversible CBA/H-T6 chimaeras. We suggest that with tolerance to the leukaemia virus being maintained and in the absence of anti-viral antigenic complexes, tumour-specific sites can be recognized and thus tumours are eliminated. This hypothesis remains to be proven. Images Fig. 1 Fig. 2