A novel approach to sexing bovine blastocysts using male-specific gene expression.

When examining gene expression profiles for the purposes of assessing embryo quality, it is imperative that sex be considered, because many embryonic transcripts have sex-related expression patterns. The objective of this study was to systematically examine eight Y chromosome linked genes (DDX3Y, EIF1AY, HSFY, SRY, TSPY, USP9Y, ZFY, and ZRSR2Y) to characterize their expression in bovine blastocysts and to examine the usefulness of this expression for the purpose of RNA-based embryo sexing. In order to examine the expression of these genes, pools of blastocysts (groups of 10 and 20) as well as single embryos (N = 50) were analyzed with reverse transcriptase polymerase chain reaction (RT-PCR) and reverse transcriptase quantitative PCR (RT-qPCR). Of the 50 single embryos, 32 were concurrently sexed with DNA-based methods. Transcripts of DDX3Y, EIF1AY, TSPY, USP9Y, ZFY and ZRSR2Y were detected in the pooled and single blastocysts, but no transcripts were detected for HSFY or SRY. After performing DNA-based sexing experiments, we concluded that this expression was restricted to the male embryos. The consistency of the expression varied according to the gene as well as the specific primer set. Three genes were expressed in the full set of male embryos, DDX3Y, USP9Y, and ZRSR2Y and therefore represent good candidates for RNA-based sexing methods.

Dependent patients discharged home from PRM departments: relevant indicators.

OBJECTIVE: To evaluate simple, measurable indicators of optimal organizational procedures for the hospital-to-home discharge of dependent patients. MATERIAL AND METHOD: All the general practitioners (GPs) in the Maine-et-Loire county of France were sent a questionnaire asking them to rank the three main criteria (from the most important to the least significant) from a list of 14. We analyzed the median ranking for each item and identified the most important items in terms of their relative frequency. RESULTS: The response rate was 10.77% (104 out of 966). Four criteria had a median score over 9: contact with the GP prior to discharge, informing the GP of the discharge date, training for the patient and his/her family in activities of daily living and providing a list of people to be contacted in the event of a problem at home. Respite hospitalization (in the event of difficulties at home) was cited as one of the three most relevant criteria. DISCUSSION-CONCLUSION: The criteria highlighted by the GPs were not highly specific for the discharge of a dependent patient. However, it would be interesting to extend this study by interviewing other stakeholders and determining whether these criteria indeed improve the organization of hospital-to-home discharge.

Normal retina releases a diffusible factor stimulating cone survival in the retinal degeneration mouse.

The role of cellular interactions in the mechanism of secondary cone photoreceptor degeneration in inherited retinal degenerations in which the mutation specifically affects rod photoreceptors was studied. We developed an organ culture model of whole retinas from 5-week-old mice carrying the retinal degeneration mutation, which at this age contain few remaining rods and numerous surviving cones cocultured with primary cultures of mixed cells from postnatal day 8 normal-sighted mice (C57BL/6) retinas or retinal explants from normal (C57BL/6) or dystrophic (C3H/He) 5-week-old mice. After 7 days, the numbers of residual cone photoreceptors were quantified after specific peanut lectin or anti-arrestin antibody labeling by using an unbiased stereological approach. Examination of organ cultured retinas revealed significantly greater numbers of surviving cones (15-20%) if cultured in the presence of retinas containing normal rods as compared with controls or cocultures with rod-deprived retinas. These data indicate the existence of a diffusible trophic factor released from retinas containing rod cells and acting on retinas in which only cones are present. Because cones are responsible for high acuity and color vision, such data could have important implications not only for eventual therapeutic approaches to human retinal degenerations but also to define interactions between retinal photoreceptor types.

Photoreceptor transplants increase host cone survival in the retinal degeneration (rd) mouse.

Retinal transplants offer a potentially interesting approach to treating human retinal degenerations, but so far little quantitative data are available on possible beneficial effects. We isolated photoreceptor layers from normal-sighted mice and grafted them into the subretinal space of retinal degeneration (rd) mice lacking rod photoreceptors. At 2 weeks after surgery, the numbers of residual host cone photoreceptors outside the graft zone were quantified following specific labelling. Examination of operated retinas revealed highly significantly greater numbers of surviving cones (mean of 38% more at 2 weeks) within the central field compared to sham-operated paired control retinas (p < 0.01). These are the first quantified data indicating a trophic effect of transplanted photoreceptors upon host cone cells. As cone cells are responsible for high acuity and colour vision, such data could have important implications not only for eventual therapeutic approaches to human retinal degenerations but also to understanding underlying interactions between retinal photoreceptors.

[Retinal grafts: biological problems and clinical stakes]. / Greffes de rétine: problèmes biologiques et enjeux cliniques.

Retinal transplantation, formerly perceived as unrealistic, has become over the past decade a major clinical and biological undertaking in several laboratories and eye clinics. We describe the insights gained through the pioneering experimental works of Del Cerro et al, Turner et al, Gouras et al, Aramant et al, Lund et al e.g. the survival of transplants, the lack of immune response to photoreceptors, their integration and expression of neuronal markers, but also the dysplastic arrangement into rosettes and the lack of a definitive proof for functionality. Our laboratory has undertaken to establish the trophic and synaptic functions of sheets of photoreceptors transplanted, as described by Silverman et al, in the subretinal space of mutant rd mice carrying a retinal degeneration similar to human retinitis pigmentosa. Clinical applications to this condition as well as in cases of end-stage age related macular degeneration are discussed.

[Influence of visible interferences on biochemical assays realised on Au 5231, AU 5223 (Olympus) and CL 7200 (Shimadzu)]. / Influence des interférences visibles sur les dosages de biochimie réalisés sur AU 5231, AU 5223 (Olympus) et CL 7200 (Shimadzu).

The impact of the major interferents (hemolysis, bilirubin, turbidity), on the quality of biochemical tests, was evaluated on multiparametric analysers (CL 7200 Shimadzu, Japan/Ciba-Corning, France; AU 5231 and AU 5223 Olympus, Japan/bioMérieux, France), according to the SFBC instructions. Interferences were detected in 33 cases upon 165 tests realized, that is to say 20% of the performed analysis. Turbidity was the most frequent cause of interference (7.8%), followed by hemolysis (8.5%) and bilirubin (3.6%). The use of a sample blank, a bireagent, the change of reagent, the change of the secondary wavelength or the modification of the measurement times, allowed us to reduce more than 80% of the interferences. Only three interferences remained: hemolysis upon the measurement of TGO and potassium, and bilirubin upon the measurement of creatinine. For these parameters, a suitable note using the Olympus factors (semi quantitative expression of the importance of the three interferents) is reported on the answer sheet.

[In vivo sensitivity of Plasmodium falciparum to amodiaquine in the town of Edea (Cameroon)]. / Sensibilité in vivo de Plasmodium falciparum à l'amodiaquine dans laville d'Edéa (Cameroun).

In the town of Edea, where falciparum malaria is hyperendemic, an in vivo study of amodiaquine sensitivity of the local strain of the parasite is performed in school children 6 to 12 years of age: 184 children with parasites in their blood and no chloroquine in their urine are given amodiaquine orally. In 96 children having taken 35 mg per kg body weight of amodiaquine base, none is any longer harboring parasites in his blood as soon as the second day following the end of the 3 days treatment (15 mg/kg body weight the first day, 12 mg/kg the second day and 8 mg/kg the third day). The 73 children having taken 27 mg/kg body weight of amodiaquine base are cleared of their parasites at the rate of 93% on the seventh day of the experiment. The 15 children having only taken 15 mg/kg body weight of amodiaquine base are cleared of their parasites at the rate of 54%. At the utilized doses, amodiaquine is generally well tolerated. Conjunctival hyperhaemia has only been observed as a secondary effect in 9 children among the 184 children treated but this side symptom disappeared soon after the end of the treatment. Aminotransferases blood levels are not modified in the course of the amodiaquine treatment.

[Comparative study of the malaria indices in Nanga-Eboko, Yaounde and Edea (Cameroon)]. / Etude comparée des indices paludométriques à Nanga-Eboko, Yaoundé et Edéa (Cameroun).

The authors report the results of sample surveys carried out in september 1986 in Yaounde and Nanga-Eboko and in january 1987 in Edea (Cameroon) to evaluate the main malarial indexes among 0-15 years old children. These investigations suggest that malaria is hypoendemic in Yaounde and mesoendemic in Nanga-Eboko during the rainy months. In Edea malaria is hyperendemic.

Detection with a monoclonal antibody of an antigen, characteristic of the genus Schistosoma, excreted in the urine.

In the host, the antigen excreted by schistosomes in the circulating blood is concentrated in the urine. A mouse monoclonal antibody of the IgM class type lambda, directed against an epitope of the intestinal epithelium of the adult worm, is obtained. The antigen found in the urine of the host as well as the monoclonal antibody has been previously characterized. It is of a polysaccharidic nature, is thermostable and specific for the genus Schistosoma. The antigen is found at all stages of the life cycle and, particularly, in the egg where it is found in large amounts. Detection of the antigen is by means of inhibition of the passive haemagglutination test. There is a fundamental advantage in detecting the metabolic antigen excreted by schistosomes instead of looking for circulating antibodies. The antigen is directly released by the parasite itself, antibodies being, by contrast, produced by the host, indirectly therefore, and in a way that varies from one individual to the next. Collecting urine specimens is, for field workers, easier than obtaining blood from the inhabitants. The detection of the antigen in the urine is made a rather simple procedure since the antigen is concentrated by the kidney and free in urine, instead of remaining conjugated with antibodies like it is in the blood. When used in the Cameroon for the study of prevalence in two foci of schistosomiasis, intestinal (Nalassi Emana) and urinary (Barombi Kotto), the test detecting the antigen in urine gives good correlations with the parasitological examinations looking for eggs of S. mansoni and S. haematobium in feces and urine.

[Epidemiological aspects of malaria in 2 villages of the Manyemen forest region (Cameroon, southwest province)]. / Aspects épidémiologiques du paludisme dans deux villages de la région forestière de Manyemen (Cameroun-province du Sud-ouest).

In terms of parasitaemia in blood, the prevalence was 18.7% for Plasmodium falciparum, 10.5% for P. ovale and 1% for P. malariae in the villages of Mungo Ndor and Kokobuma. The plasmodial index of children from 2 to 9 years of age was 38.2% and the spleen index 26.6%, with a mean HACKETT score of 1.56. Malaria is thus meso-endemic in the region. The immunofluorescent test performed with a P. falciparum antigen was positive in 66% of the children in the same age group. Parasites were seen in 15% of new-born children. The malaria infection indices were higher in Mungo Ndor, which is located on the main road in the vicinity of the Manyemen hospital, than in Kokobuma. Morbidity and mortality due to malaria remain major problems in this part of the South-West Province, in spite of improving primary health care facilities and free chloroquine distribution. Anopheles gambiae is the main vector of malaria in the area, and transmission is interrupted only during the short dry season.

[A fatal case of chloroquine-resistant neuromalaria in a traveller to Cameroon]. / A propos d'un cas fatal de neuropaludisme chloroquinorésistant chez un voyageur au Cameroun.

A death from chloroquinoresistant falciparum malaria contracted by a traveller under chloroquine chemoprophylaxis during a stay in the Cameroon provides the authors with an opportunity to analyze the extension of chloroquinoresistance in Africa. Having appeared there about 10 years ago, it remained for a long time localized in East Africa. In recent years, it has moved West. Today, 17 countries located from the North to the South between Sudan and Mozambique, and from East to West between Kenya and the Cameroon are affected, as are the Malagasy Republic, the Comores and Zanzibar. The strains resistant to chloroquine are still sensitive to amodiaquine. In certain regions which are widespread in the continent, resistance to pyrimethamine has become evident in the last 30 years; resistance to pyrimethamine-sulfadoxine and pyrimethamine-dapsone has appeared in the last 3 years in East Africa. In the light of the complexity of the problem of malaria chemoprophylaxis, the authors emphasize the necessity of resensitizing the public at large with respect to personal protection against biting anopheline mosquitos, especially during the hours of darkness. The medical profession should equally be made aware of these new problems. Three types of chemoprophylaxis in relation to the areas visited are proposed: the first for West Africa; the second for Central Africa, neighbouring regions and the Malagasy Republic and the Comores; the third for the 3 East Africa countries the most affected, which are Kenya, Burundi and Tanzania.

[Epidemiology of filariasis (onchocerciasis and bancroftosis) in the Tala-Mokolo region (Mandara Mountains of North Cameroon)]. / Epidémiologie des filarioses (onchocercose et bancroftose) dans la région de Tala-Mokolo (Monts Mandara-Nord Cameroun).

The effect of the bancroftian filariasis on the health of the inhabitants are minimal in the study area, where the prevalence of the disease (nocturnal microfilaremia) is 0.4% only. But skin snips harboring Onchocerca microfilariae are seen in 12.1% of the sample studied. Onchocerciasis skin lesions, low visual acuity and blindness are often seen in the villages where the prevalence is high, among inhabitants more than 40 years of age. Simulium damnosum is seldom seen in the area, except in the large streams and rivers like the mayo Tsanaga and the mayo Djingliya. Larvae and nymphae of this species hardly succeed in developing on the overspelling of the small dams, this being due more to a discontinuous run of the water in the overspilling than to a to high speed of the water.

[Urban and rural malaria in Niger. The case of the Department of Maradi]. / Paludisme urbain et rural au Niger. Le cas du département de Maradi.

In Niger, two communities, a urban one (Maradi) and a rural one (Tarna), living in a same malarious Soudano-Sahelian climatic area, were compared during wet and dry seasons by different methods (spleen rate, parasite rate, parasite count, level of immunity). During wet and dry season, spleen rates (0.7% and 2.6%) and parasite rates (1% and 1.6%) are considerably lower at Maradi than those observed at Tarna (spleen rates: 38% and 42%; parasites rates: 10% and 22.5%). It is shown that seroconversion occurs later in urban sector (100% between 20 and 29 year age) than in rural sector (100% between 10 and 15 year age). All malarious indicators were negative in a high percentage of Maradi-born children under 9 (79% during wet season and 81% during dry season) against respectively 28% and 31% at Tarna. Better paludometric index in urban community is a favourable sanitary indication. But epidemiological control must be prosecuted since the number of non-immune subjects is increased.

[Evaluation of the clinical, parasitological and immunological indices of malaria in the Bonny's Bay area of Central Africa]. / Evaluation des indices cliniques, parasitologiques et immunologiques du paludisme dans la région de la Baie de Bonny en Afrique Centrale.

The authors report the results of 4 sample surveys carried out in Equatorial Guinea and Cameroon in the Bonny's Bay area, to evaluate the main malarial indexes. In rural and semi-rural areas one notice a high rate of fever attacks and splenomegaly. Parasitaemia is more important in rural areas than in urban areas and is decreasing where the therapeutic pressure is strong and long-lasting. Immunological indexes are high everywhere.