Prevalence and genetic variation of salivary gland hypertrophy virus in wild populations of the tsetse fly Glossina pallidipes from southern and eastern Africa.

The Glossina pallidipes salivary gland hypertrophy virus (GpSGHV) is a rod-shaped, non-occluded double-stranded DNA virus that causes salivary gland hypertrophy (SGH) and reduced fecundity in the tsetse fly G. pallidipes. High GpSGHV prevalence (up to 80%) makes it impossible to mass-rear G. pallidipes colonies for the sterile insect technique (SIT). To evaluate the feasibility of molecular-based GpSGHV management strategies, we investigated the prevalence and genetic diversity of GpSGHV in wild populations of G. pallidipes collected from ten geographical locations in eastern and southern Africa. Virus diversity was examined using a total sequence of 1497 nucleotides (≈ 1% of the GpSGHV genome) from five putative conserved ORFs, p74, pif1, pif2, pif3 and dnapol. Overall, 34.08% of the analyzed flies (n=1972) tested positive by nested PCR. GpSGHV prevalence varied from 2% to 100% from one location to another but phylogenetic and gene genealogy analyses using concatenated sequences of the five putative ORFs revealed low virus diversity. Although no correlation of the virus diversity to geographical locations was detected, the GpSGHV haplotypes could be assigned to one of two distinct clades. The reference (Tororo) haplotype was the most widely distributed, and was shared by 47 individuals in seven of the 11 locations. The Ethiopian haplotypes were restricted to one clade, and showed the highest divergence (with 14-16 single nucleotide mutation steps) from the reference haplotype. The current study suggests that the proposed molecular-based virus management strategies have a good prospect of working throughout eastern and southern Africa due to the low diversity of the GpSGHV strains.

Susceptibility of North-American and European crickets to Acheta domesticus densovirus (AdDNV) and associated epizootics.

The European house cricket, Acheta domesticus L., is highly susceptible to A. domesticus densovirus (AdDNV). Commercial rearings of crickets in Europe are frequently decimated by this pathogen. Mortality was predominant in the last larval stage and young adults. Infected A. domesticus were smaller, less active, did not jump as high, and the adult females seldom lived more than 10-14 days. The most obvious pathological change was the completely empty digestive caecae. Infected tissues included adipose tissue, midgut, epidermis, and Malpighian tubules. Sudden AdDNV epizootics have decimated commercial mass rearings in widely separated parts of North America since the autumn of 2009. Facilities that are producing disease-free crickets have avoided the importation of crickets and other non-cricket species (or nonliving material). Five isolates from different areas in North America contained identical sequences as did AdDNV present in non-cricket species collected from these facilities. The North American AdDNVs differed slightly from sequences of European AdDNV isolates obtained in 1977, 2004, 2006, 2007 and 2009 and an American isolate from 1988. The substitution rate of the 1977 AdDNV 5kb genome was about two nucleotides per year, about half of the substitutions being synonymous. The American and European AdDNV strains are estimated to have diverged in 2006. The lepidopterans Spodoptera littoralis and Galleria mellonella could not be infected with AdDNV. The Jamaican cricket, Gryllus assimilis, and the European field cricket, Gryllus bimaculatus, were also found to be resistant to AdDNV.

Hytrosaviridae: a proposal for classification and nomenclature of a new insect virus family.

Salivary gland hypertrophy viruses (SGHVs) have been identified from different dipteran species, such as the tsetse fly Glossina pallidipes (GpSGHV), the housefly Musca domestica (MdSGHV) and the narcissus bulbfly Merodon equestris (MeSGHV). These viruses share the following characteristics: (i) they produce non-occluded, enveloped, rod-shaped virions that measure 500-1,000 nm in length and 50-100 nm in diameter; (ii) they possess a large circular double-stranded DNA (dsDNA) genome ranging in size from 120 to 190 kbp and having G + C ratios ranging from 28 to 44%; (iii) they cause overt salivary gland hypertrophy (SGH) symptoms in dipteran adults and partial to complete sterility. The available information on the complete genome sequence of GpSGHV and MdSGHV indicates significant co-linearity between the two viral genomes, whereas no co-linearity was observed with baculoviruses, ascoviruses, entomopoxviruses, iridoviruses and nudiviruses, other large invertebrate DNA viruses. The DNA polymerases encoded by the SGHVs are of the type B and closely related, but they are phylogenetically distant from DNA polymerases encoded by other large dsDNA viruses. The great majority of SGHV ORFs could not be assigned by sequence comparison. Phylogenetic analysis of conserved genes clustered both SGHVs, but distantly from the nudiviruses and baculoviruses. On the basis of the available morphological, (patho)biological, genomic and phylogenetic data, we propose that the two viruses are members of a new virus family named Hytrosaviridae. This proposed family currently comprises two unassigned species, G. pallidipes salivary gland hypertrophy virus and M. domestica salivary gland hypertrophy virus, and a tentative unassigned species, M. equestris salivary gland hypertrophy virus. Here, we present the characteristics and the justification for establishing this new virus family.

A new densovirus isolated from the mosquito Culex pipiens (Diptera: culicidae).

We have isolated from a laboratory strain of Culex pipiens in which abnormal larval mortality occurred a small icosahedral nonenveloped DNA virus sharing the main biological and biophysical properties of densoviruses (DNVs). Unlike DNVs isolated previously from Aedes species, i.e. the AaeDNV and the AalDNV (Afanasiev, B.N., Galyov, E. E., Buchatsky, L.P., Kozlov, Y.V., 1991. Nucleotide sequence and genomic organization of Aedes densonucleosis virus. Virology 185, 323-336; Boublik, Y., Jousset, F.-X., Bergoin, M., 1994. Complete nucleotide sequence and genomic organization of the Aedes albopictus Parvovirus (AaPV) pathogenic for Aedes aegypti larvae. Virology 200, 752-763), this mosquito DNV named CpDNV possesses a genome of 6 kb separately encapsidated in stoichiometric proportion as 'plus' and 'minus' strands. The lack of sequence homology between the CpDNV and the AalDNV genome and of antigenic cross-reactivity between their capsid polypeptides indicates that these two mosquito DNVs are phylogenetically distant. In contrast, the CpDNV appears to be related to the Junonia coenia densovirus (JcDNV) at both serological and genomic levels.

The spheroidin of an entomopoxvirus isolated from the grasshopper Anacridium aegyptium (AaEPV) shares low homology with spheroidins from lepidopteran or coleopteran EPVs.

Based on virion morphology, the current virus taxonomy groups entomopoxviruses (EPVs) (Poxvirus: Entomopoxvirinae) from coleopteran and dipteran hosts in separated genera, wilts it keeps viruses infecting either lepidopteran or orthopteran hosts in the same genus. In contrast to the morphological criteria, the few data available from recent studies at the genetic level have suggested that EPVs infecting different insect orders are phylogenetically distant. In order to elucidate EPVs phylogeny we have cloned and sequence the highly conserved/highly expressed spheroidin gene of Anacridium aegyptium entomopoxvirus (AaEPV). This gene and its promoter is of interest for the development of genetic engineering on EPVs. The spheroidin gene was located in the AaEPV genome by Southern blot and hybridisation with specific degenerated oligonucleotides probes synthesised after partial sequencing of the purified spheroidin protein. A total of 3489 bp were sequenced. This sequence included the coding and promoter region of 969 residues 108. 8 kDa protein identified as spheroidin. AaEPV spheroidin contains 21 cysteine residues (2.2%) and 14 N-glycosylation putative sites distributed along the sequence. The cysteine residues are particularly abundant at the C-terminal end of the protein, with 11 residues in the last 118 aa. Our results confirm that the spheroidin is highly conserved only between EPVs isolated from the same insect order. Polyclonal antibodies raised against AaEPV spherules specifically revealed spheroidin in Western Blots failing to cross-react with MmEPV or AmEPV spheroidins or MmEPV fusolin. Comparison of spheroidins at the aa level demonstrate that AaEPV spheroidin shares only 22.2 and 21.9% identity with the lepidopteran AmEPV and the coleopteran MmEPV spheroidins, respectively, but 82.8% identity with the orthopteran MsEPV spheroidin. Only two highly conserved domains containing the sequence (V/Y)NADTG(C/L) and LFAR(I/A) have been identified in all known spheroidins. The phylogenetic tree constructed according to the CLUSTALX analysis program revealed that EPVs are clearly separated in three groups - lepidopteran, coleopteran and orthopteran - according to the insect order of the virus hosts. In base to our results, the split of the genus Entomopoxvirus B dissociating lepidopteran and orthopteran EPVs into two different genera is suggested.

Venereal and vertical transmission of the Aedes albopictus parvovirus in Aedes aegypti mosquitoes.

Following per oral infection of Aedes aegypti larvae with Aedes albopictus parvovirus (AaPV), infected males and females adults were tested for their ability to transmit the virus venereally and vertically, respectively. Both types of transmission were observed. A low percentage (2.2%) of AaPV-free females were found contaminated by the virus after mating with AaPV-infected males. Although no significant difference was observed in the fecundity of orally infected and virus-free females, 17.1% of infected ones died before egg laying, whereas no mortality occurred during the same period in virus-free females. There was a clear relationship between the virus titer in the orally infected females and both mortality and infection in their offspring. The virus titer averaged 10(6.2) 50% tissue culture infective doses (TCID50s) in F1 females and 10(3.3) TCID50 in F1 females. Nevertheless, AaPV did not persist in an experimentally infected population of mosquitoes beyond the second generation.

A titration procedure of the Junonia coenia densovirus and quantitation of transfection by its cloned genomic DNA in four lepidopteran cell lines.

A sensitive and reproducible tissue culture biossay method was developed based on indirect immunofluorescence to titrate virus suspensions of the Junonia coenia densovirus (JcDNV) and to quantify transfections by its cloned genomic DNA. Four lepidopteran cell lines, the SPC-SL 52 from Spodoptera littoralis, the SPC-PL 40 and the SPC-PL 65 cells derived from Spodoptera litura ovaries and hemocytes, respectively, and the SC-LD 135 from Lymantria dispar were compared for their efficiency to support viral replication. The viral titres expressed as TCID50/ml averaged 10(5) for SPC-SL 52, SPC-PL 40 and SC-LD 135 cells, but were above 10(7) for SPC-PL 65 cells. Even with this most sensitive cell line, the rate of infected cells did not exceed 75% and decreased progressively by serial subcultures. Two transfection protocols were used to compare the sensitivity of the same four cell lines to a recombinant plasmid encompassing an infectious sequence of JcDNV genome. SPC-SL 52 cells were found to be the most sensitive, and the lipofection method resulted in about a 5-fold increase compared to the calcium phosphate precipitation protocol. The rescued virions proved to be infectious and the restriction profiles of their DNA were identical to that of wild type virions.

Expression of heterologous promoters in Lutzomyia longipalpis and Phlebotomus papatasi (Diptera: Psychodidae) cell lines.

To establish a transient expression system for genes introduced into sand fly cell lines, we tested the expression of the luciferase reporter gene under control of different promoters. Towards this end, we lipofected cell lines obtained from New and Old World sand flies, LL-5 from Lutzomyia longipalpis Lutz & Neiva and PP-9 from Phlebotomus papatasi Scopoli, respectively. The relative levels of luciferase expression were studied under control of Drosophila melanogaster Meigen heat shock protein 70 (hsp70), human cytomegalovirus, simian virus 40 or Junonia coenia (Hübner) densovirus (P9) promoters. The Drosophila heat shock protein 70 promoter, originating from insect genes, functioned as a strong promoter in both cell lines. Promoters from the different virus genes also were capable of driving transgene expression in both cell lines.

Treatment of a case of congenital pseudarthrosis of the tibia-fibular osteosynthesis.

We report the results of a case of congenital pseudarthrosis of the tibia treated by tibiofibular synthesis. A 1-year old girl was first treated by intramedullary fixation followed by an intertibiofibular bone graft. This method failed. She then underwent a new operation that was associated two simultaneous approaches, correction of the axis, tibiofibular synthesis and a new intertibiofibular bone graft. Union was achieved four months later. The child has now been followed up for 20 years. During this time, she has led a normal life as we have observed a "tibialisation" of the fibula. Intramedullary fixation has a success rate of 75% but requires repetitive insertion of intramedullary or telescopic rods. Transplantar intramedullary rods are responsible for significant ankle stiffness. Tibiofibular synthesis associated with an intertibiofibular bone graft after correction of the axis is the equivalent of vascularised graft of the fibula but with neither the difficulties of microsurgery nor valgus deformities of the ankle.

The treatment of idiopathic scoliosis in adolescents: rotation or in situ bending?

Rotation alone is not fully efficient in order to correct all types of scoliotic curvatures. We report a series of 30 cases instrumented with the EUROS spine system and analyse reductions obtained with in situ rotation or bending alone or with combined maneuvres. The average age of surgery is 17 years for this series composed of 24 female and 6 male patients. The average follow-up is 2.3 years. The curve patterns are displayed with 6 major thoracic, 5 genuine double major, 4 double major thoracic predominant, 6 double major lumbar predominant and 9 double thoracic curves. Combination of both reduction techniques is advisable and is to be made according to the type of curvature and its reducibility in situ bending is made easier with this system without lockers and by reduced diameter of the rod.

The choice of treatment for simple bone cysts of the upper third of the femur in children.

A retrospective study of a series is presented of 19 simple cysts of the upper third of the femur in children of a mean age of 7 years. The patients included 4 adolescents and 15 children under the age of 10. Mean recurrence was 6 years. Different treatment methods were utilized: - surgical treatment: curettage-bone grafting with or without osteosynthesis, - intra-cystic corticoid injection, - orthopedic treatment. Cysts may be small with a low lytic potential, in this case intra-cystic corticoid injections are very effective; or bigger in size with a risk of fracture or have already caused a fracture, the treatment is then surgical but should always be combined with osteosynthesis.

X-ray transparency interpositions after reduction of traumatic dislocations of the hip in children.

Traumatic dislocations of the hip are much less frequent in children than in adults. If some complications are well known (avascular necrosis of the femoral head, recurrent dislocation), the lateralization of the head by x-ray transparency interposition is less appreciated. We report 4 cases of x-ray transparency interposition in 15 reductions of incomplete traumatic dislocation of the hip. These 4 dislocations were posterior in non-pathological hips, three on the left side and one on the right. Our reductions have been easy and the hips remained stable. However, the post-reduction roentgenogram shows the same unusual enlarged joint space corresponding to the lateralization of the femoral head. Four arthrotomies were performed, tracing a cartilaginous fragment free or fixed to the articular capsule into the femoral joint. We have found a concentric reduction of the hip in 4 cases and enlargement from 1 to 3 mm of the joint space of the hip. After several years, articular function of the four hips is normal, nevertheless, they have shown a coxa valga with a femoral elongation to 10 mm, a coxa magna with an articular space wider than 1 mm, 2 very good concentric reductions of the hip. The lateralization of the head after reduction had to be detected as soon as the first radiographic control because it can cause immediate instability or arthrosis of the hip. Because of the cartilaginous or capsular composition and the size of the interposition, an arthrotomy is performed in front of a radiological enlarging of the space joint, or a loss of parallelism of osteochondral acetabulum bone. Computerized tomography and MRI can reveal the nature, the place of the included fragment, as well as the surgical approach. An arthrogram is not absolutely indispensible. Surgical exploration should avoid degenerative arthrosis after a lateralisation left untreated.

Anterior approach versus posterior approach to surgical treatment of children's supracondylar fractures: comparative study of thirty cases in each series.

The purpose of this study was to consider the surgical treatment of severe supracondylar fractures of the elbow in children, and to compare the anterior approach with the posterior approach used in two homogeneous groups of 30 cases each by two experienced surgeons. Control procedures were maintained with the children of both groups when the plaster was removed, during the fourth month after surgery, and throughout the follow-up that continued for more than 1 year. A posterior approach to surgery is simpler than an anterior approach, but it creates supplementary anatomic damage that can cause circulatory disorders in the external condyle and a higher percentage of limitation in articulation mobility. Thus, although the anterior approach is more technically demanding, it gives better functional results. Because this approach concerns a zone already damaged by the trauma, it eliminates hematoma in the anterior brachial muscle and again places the fragments in the untouched shell of the periosteum.

[Disk calcifications in children. Apropos of 17 cases]. / Calcifications discales de l'enfant. A propos de 17 cas.

Seventeen cases of calcification of the intervertebral discs in childhood have been seen. Only cervical lesions gave symptoms. In several cases the lesions were multifocal (cervical and thoracic) and there was a familial history in one case. Two cases were observed for 10 and 8 years respectively. It was noted that cervical calcifications tend to disappear while thoracic ones persist but are asymptomatic. The authors postulate that this is due to differences in the physiology of the nucleus and in the biomechanics at different levels of the spine. A high frequency of associated spinal deformities was noted. For this reason, the late prognosis must be guarded.

[Resection of hemivertebrae. A review of seven cases (author's transl)]. / Technique sûre d'exérèse chirurgicale des hémi-vertèbres.

The technique used by the authors for removal of hemivertebrae is fully described. Two approaches are made, one posterior and one lateral at the same surgical session. This allows satisfactory control of roots, medulla and radicular vessels. The indications are described with reference to the age of the patient, the level of the lesion and the type of anomaly. It is concluded that it is a valuable procedure in progressive severe congenital scoliosis. 7 cases are reported without any neurological complication.

[Surgical treatment of the rupture of the inferior tendon of the biceps. Apropos of 6 cases]. / Traitement chirurgical des ruptures du tendon distal du biceps brachial. A propos de six observations.

INTRODUCTION: The purpose of this study is to compare the results of direct reinsertion of the distal bicipital tendon to the bicipital tuberosity, with other methods described in the literature. MATERIAL AND METHOD: This study is based on six athletic patients, all male, ranging in age from 42 to 62 years old. The rupture of the biceps occurred during an unusually violent effort in flexion and forced supination of the arm at a 90 degree of flexion. The surgical procedure which took place between a week and a month after the accident, consisted in a direct reattachment of the detached tendon. RESULTS: No paralysis in the area of the radial nerve occurred. All patients regained normal range of motion of the joints with maintained strength. DISCUSSION: Faced with a recent lesion in a young patient, a reinsertion on the bicipital tuberosity is desirable in order to maximize the strength and movement of the biceps. This simple technique uses a single approach. It does not weaken the bicipital tuberosity nor the muscular function of the biceps.

[Tenotomy of the psoas muscle in slow reduction of congenital hip dislocations]. / La ténotomie du psoas au cours de la réduction lente des luxations congénitales de hanche.

PURPOSE OF THE STUDY: The aim of this study was to relate the efficacy of cutting the psoas tendon in case of high congenital hip dislocation treated by slow reduction according to Somerville-Petit. MATERIAL: Twenty psoas tenotomies were realised between 1982 and 1992. The average age was 11 months 15 days old, they were 15 girls and 5 boys. Among them, only 9 patients had never been treated. METHOD: By a short surgical approach close to the lateral lip of Scarpa's triangle, we cut the psoas tendon at the musculotendinous junction doing this suppresses the obstacle that interferes with the femoral head coming down. RESULTS: Results have been evaluated on clinically and especially radiologically. All hips remain stable except one case. A graduated subluxation of the hip occured after removal of the plaster. 18 hips have been revised between the third and fifth year. Clinical results were excellent, on radiological controls 5 hips were considered flawless, 11 satisfying, and 2 were imperfect. Only 2 children have been seen ten years later with excellent clinical and radiological results. DISCUSSION: The hypertrophic psoas tendon interposes between the femoral head and the acetabulum creasing the capsule and the limbus. Cutting tendon of the psoas will decrease the numerous unexplained failures that occured during slow reduction according to Sommerville-Petit method. The use of these surgical techniques may make more outstanding the tenotomies time. CONCLUSION: Tenotomy of the psoas tendon realized by a short surgical approach without any articular effraction, allowed us to obtain complete reduction of the hips dislocation in these 20 patients.

Viruses as nanoparticles: structure versus collective dynamics.

In order to test the application of the "nanoparticle" concept to viruses in terms of low-frequency dynamics, large viruses (140-190 nm) were compared to similar-sized polymer colloids using ultra-small-angle x-ray scattering and very-low-frequency Raman or Brillouin scattering. While both viruses and polymer colloids show comparable highly defined morphologies, with comparable abilities of forming self-assembled structures, their respective abilities to confine detectable acoustic vibrations, as expected for such monodisperse systems, differed. Possible reasons for these different behaviors are discussed.

Congenital scoliosis: a frontal plane evaluation of 251 operated patients 14 years old or older at follow-up.

INTRODUCTION: Congenital scoliosis, carrying an incidence between 0.5 and 1 per 1000 births, raise the problem of their evolutive potential. HYPOTHESIS: Some predictive factors for the evolution of scoliotic curvature due to congenital vertebral malformation (CVM) can be found. MATERIAL AND METHODS: This was a retrospective multicenter study of 251 patients, at least 14 years old when evaluated at end of follow-up, with CVM and spinal deformity predominating in the frontal plane. RESULTS: 38.8% of patients showed associated neurologic, visceral or orthopedic abnormalities. CVM was single in 60.6%, double in 20.3%, triple in 6.4% and multiple in 12.7% of cases. 34.1% of CVMs were thoracic. Congenital scoliosis curvature was single in 88.8% of patients, double in 10% and triple in 1.2%. Mean curvature angle was 31.7° at diagnosis (range, 0-105°) and 41.3° preoperatively (range, 10-105°). Sixty-one patients showed associated kyphosis. Mean change in postoperative curvature angle over follow-up was 1.6° (range, -20° to 38°) in the 73 patients managed by arthrodesis, -0.4° (-24° to 30°) in the 64 managed by epiphysiodesis, and 0.4° (-18° to 35°) in the 49 managed by hemivertebral (HV) resection. Results were found to correlate significantly with age at surgery for patients managed by epiphysiodesis, but not for those managed by HV resection or arthrodesis. DISCUSSION: More than 30% of congenital scolioses involve associated intraspinal abnormality. All CVM patients should therefore undergo medullary and spinal MRI to assess the CVM in all three planes, and the medullary canal and its content. The evolution of scoliotic curvature induced by CVM is hard to predict. Several factors are to be taken into account: CVM type, number and location, and patient age. Curvature progression may be slow or very fast. It accelerates during the peak of puberty, stabilizing with bone maturity. Surgery is mandatory in evolutive scoliosis. Four procedures may be recommended, according to type of CVM and especially to patient age: arthrodesis, convex epiphysiodesis, HV resection or rib distraction. Surgery seeks to correct the spinal deformity induced by the CVM and prevent compensatory curvature and neurologic complications, while conserving sagittal and frontal spinal balance and sparing as many levels as possible. In case of HV involvement, the procedure of choice is CVM resection, which provides 87.5% good results in this indication; the procedure is relatively safe, conservative of spinal levels, and without age limit. LEVEL OF EVIDENCE: Level IV. Retrospective study.