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Here, we present the complete coding sequences of two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains that were recovered from a nasopharyngeal swab from a female patient and the second viral passage in cell culture. After testing, both strains were identified as BA.5.2.20, a subvariant of Omicron.

Here, we describe the coding-complete sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strain HM36, identified as a strain of concern of B.1.1.529+BA (Omicron).

COVID-19 caused by the virus SARS-CoV-2 has gripped essentially all countries in the world, and has infected millions and killed hundreds of thousands of people. Several innovative approaches are in development to restrain the spread of SARS-CoV-2. In particular, BCG, a vaccine against tuberculosis (TB), is being considered as an alternative therapeutic modality. BCG vaccine is known to induce both humoral and adaptive immunities, thereby activating both nonspecific and cross-reactive immune responses in the host, which combined could effectively resist other pathogens including SARS-CoV-2. Notably, some studies have revealed that SARS-CoV-2 infectivity, case positivity, and mortality rate have been higher in countries that have not adopted BCG vaccination than in countries that have done so. This review presents an overview of the concepts underlying BCG vaccination and its nonspecific immuological effects and protection, resulting in ‘trained immunity’ and potential utility for resisting COVID-19.

We report here the complete genome sequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains obtained from Moroccan patients with COVID-19. The analysis of these sequences indicates that the identified strains belong to the AY.33 sublineage of the Delta variant.

A new series of nucleoside derivatives was prepared from the reaction of 4-aminoantipyrene with different sugar moieties. In addition, ampyrone’s reaction with different aromatic aldehydes gave the corresponding Schiff base derivatives, which were also synthesized. Both molecular docking and in vitro antiviral activities at different concentrations of different synthesized compounds against SARS-CoV-2 were screened. All newly synthesized compounds were characterized on the basis of IR, 1H NMR and 13C NMR spectral data and physical data. The compounds were screened for potential cytotoxic activities. The molecular docking analysis showed that compounds 6b, 6e, 6c, 6f and 6d exhibited relatively higher binding energies (−8.1, −8.1, −8.3, −8.4 and −8.7 kcal/mol, respectively) compared to all the other compounds. However, the different compounds did not show any promising in vitro antiviral activities against SARS-CoV-2.

BackgroundTuberculosis represents a serious public health problem and a significant diagnostic and therapeutic challenge worldwide. Molecular diagnostic techniques are crucial in the World Health Organization’s new tuberculosis control strategy.This study aims to evaluate the performance of GeneXpert MTB/RIF (Cepheid Sunnyvale, CA, United States) in diagnosis of extra-pulmonary tuberculosis then compare it’s performance in detecting Rifampicin resistance to GenoType MTBDRplus (HAIN Life Sciences, Nehren, Germany).MethodsSamples from pulmonary and/or extra-pulmonary origins were analysed in a 21 months retrospective study. Samples were sent to the bacteriology laboratory forMycobacterium tuberculosisdetection using conventional bacteriological and molecular methods (GeneXpert MTB/RIF and MTBDRplus). Sensitivity and specificity were calculated for the stained smear and GeneXpert according to culture (Gold Standard) as well as for GeneXpert MTB/RIF in both negative and positive microscopy tuberculosis cases. Data’s statistical analysis was performed with SPSS13.0 software.ResultsSeven hundred fourteen patients’ samples were analysed; the average age was 47.21 ± 19.98 years with a male predominance (66.4%). Out of 714 samples: 285 were from pulmonary and 429 were from extra-pulmonary origins. The positivity rates for microscopy, GeneXpert MTB/RIF and culture were 12.88, 20.59 and 15.82%, respectively. These rates were 18.9, 23.85 and 20.35% for pulmonary samples and 9.71, 18.41 and 12.82% for extra-pulmonary samples, respectively. The sensitivity and specificity of GeneXpert MTB/RIF were almost the same in both pulmonary and extra-pulmonary samples (78.2 and 90.4%) and (79,3 and 90.3%) respectively.Rifampicin resistance rate found by GeneXpert MTB/RIF was 0.84%. Comparison of Rifampicin resistance obtained by GeneXpert MTB/RIF and Genotype MTBDRplus, showed 100% agreement between the two techniques for studied samples.ConclusionsThis confirms GeneXpert MTB/RIF advantage for tuberculosis diagnosis, particularly extra-pulmonary tuberculosis with negatively stained smear. The performance of GeneXpert and Genotype MTBDRplus are similar in detection of Rifampicin resistance. However, variability of detection performance according to tuberculosis endemicity deserves more attention in the choice of screening techniques of Rifampicin resistance, hence the interest of conducting comparative studies of detection performance under low and medium endemicity on large samples of tuberculosis populations.

The progression of the SARS-CoV-2 pandemic in Africa has so far been heterogeneous and the full impact is not yet well understood. Here, we describe the genomic epidemiology using a dataset of 8746 genomes from 33 African countries and two overseas territories. We show that the epidemics in most countries were initiated by importations, predominantly from Europe, which diminished following the early introduction of international travel restrictions. As the pandemic progressed, ongoing transmission in many countries and increasing mobility led to the emergence and spread within the continent of many variants of concern and interest, such as B.1.351, B.1.525, A.23.1 and C.1.1. Although distorted by low sampling numbers and blind-spots, the findings highlight that Africa must not be left behind in the global pandemic response, otherwise it could become a breeding ground for new variants.

Here, we report the identification and coding-complete genome sequences of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains obtained from patients with COVID-19. The variants identified belong to variant of concern B.1.617.2 and variant of interest B.1.617.1

INTRODUCTION: Escherichia coli is the most commonly isolated species in both community and healthcare-associated infections. Our study's purpose was to determine the rates of antibiotic resistance of E. coli isolates in hospital and community populations, track the kinetics of resistance rates of E. coli isolates to major antibiotics, particularly those prescribed for urinary tract infections and study the occurrence and evolution of multi-resistant phenotypes. METHODS: We conducted a retrospective study at the Bacteriological Department of the Mohammed V Military Hospital of Instruction, over a period of 7 years. All isolates of E. coli from inpatients and outpatients were included. Identification of bacterial isolates was based on culture, morphological and biochemical identification characteristics. Antibiotic susceptibility was studied using the Mueller Hilton agar diffusion method by using OXOID® type antibiotic discs and interpreted according to the recommendations of EUCAST/CA-SFM 2019. RESULTS: The rate of resistance of E. coli isolates to 3(rd) generation cephalosporins, imipenem and fluoroquinolones was 12%, 1% and 34%, respectively. The difference between the resistance rates of inpatient and outpatient E. coli isolates was statistically significant for most antibiotics (p

An unusual group A rotavirus (RVA) strain MAR/ma31/2011/G8P[14] was detected for the first time in Morocco in a stool sample from hospitalized child aged 18 months suffering from acute gastroenteritis and fever in 2011. Complete genome sequencing of the ma31 strain was done using the capillary sequencing technology. The analysis revealed the G8-P[14]-I2-R2-C2-M2-A11-N2-T6-E2-H3 constellation and the backbone genes: I2-R2-C2-M2-A11-N2-T6-E2-H3 are commonly found in RVA strains from artiodactyls such as cattle. The constellation was shared with another Italian zoonotic G8P[14] strains (BA01 and BA02), two Hungarian human strains (182-02 and BP1062) and a sheep RVA strain OVR762. Phylogenetic analysis of each genome segment of ma31 revealed a mixed gene configuration originated from animals and human. Comparison of the antigenic regions of VP7 and VP4 amino acid sequences between ma31 strain and selected animal and human strains bearing G8 and or P[14], showed a high level of conservation, while many substitutions was observed in comparison with RotaTeq™ and Rotarix™ vaccine strains. In contrast, alignment analysis of the four antigenic sites of VP6 revealed a high degree of conservation. These findings reveal a typical zoonotic origin of the strain and confirm a high potential for RVA zoonotic transmission between bovine and humans, allowing the generation of novel rotavirus genotypes.

Porcine astroviruses (PAstVs) have extended their distribution globally and have a high prevalence; however, their clinical significance is still under investigation. Thus far, information about their prevalence and diversity in the Indian pig population is unknown. This study is the first report on the prevalence and genetic characterization of PAstVs in diarrhoeic piglets in India. From January 2013 to December 2017, 757 samples were screened using an RT-PCR assay and PAstV infection was detected in 17.6% (133/757) pigs. Of the 133 positive samples, 79 (59.4%) were positive for PAstV alone, whereas 54 (40.6%) were found to be co-infected with porcine rotavirus A (PoRVA). Phylogenetic analysis of RdRp/capsid gene region revealed high genetic heterogeneity among PAstV sequences, with a predominance of PAstV lineage 4 and detection of lineage 2. The lineage 4 PAstVs exhibited 61.2%-94.5% sequence similarity at the nucleotide level to other reported sequences, whereas lineage 2 strain shared 66.0%-71.6% sequence identity with cognate sequences of the same lineage. This is the first report on PAstV and circulation of lineages 4 and 2 in India. Further, phylogenetic analysis indicates a multiphyletic origin of PAstV strains and suggests cross-border circulation of PAstVs with a similar genetic configuration in Asian countries.

The complete genome sequence of a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strain was obtained. The strain was isolated from a nasopharyngeal swab specimen from a female patient in Rabat, Morocco, with coronavirus disease 2019 (COVID-19). This strain belongs to clade 20A and has 12 mutations and 8 amino acid substitutions compared to the reference strain Wuhan/Hu-1/2019.

COVID-19 vaccination efficacy depends on serum levels of the neutralizing antibodies (NAs) specific to the receptor-binding domain of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein. Therefore, a high-throughput rapid assay capable of measuring the total SARS-CoV-2 NA level is urgently needed for COVID-19 serodiagnosis, convalescent plasma therapy, vaccine development, and assessment. Here, we developed a novel nanoplasmonic immunosorbent assay (NanoPISA) platform for one-step rapid quantification of SARS-CoV-2 NAs in clinical serum samples for high-throughput evaluation of COVID-19 vaccine effectiveness. The NanoPISA platform enhanced by the use of nanoporous hollow gold nanoparticle coupling was able to detect SARS-CoV-2 NAs with a limit of detection of 0.2 pM within 15 min without washing steps. The one-step NanoPISA for SARS-CoV-2 NA detection in clinical specimens yielded good results, comparable with those obtained in the gold-standard seroneutralization test and the surrogate virus-neutralizing enzyme-linked immunosorbent assay. Collectively, the one-step NanoPISA might be a rapid and high-throughput NA-quantification platform for evaluating the effectiveness of COVID-19 vaccines.

Species A rotaviruses (RVAs) are a leading cause of diarrhea in children and in the young of a large variety of mammalian and avian host species. The purpose of this study was to identify RVA in nomadic goats and calves during severe diarrhea outbreaks in 2012 and 2014 in Bouaarfa, Morocco, and to characterize the complete genomic constellation of two bovine and caprine strains (S18 and S19) and their genetic relatedness with the human strain ma31 detected in 2011 in Morocco. Partial nucleotide sequencing of VP4 and VP7 genes for the twenty-two positive samples revealed three circulating genotypes: G6P[14], G10P[14], and G10P[5] with predominance of G6P[14] genotype. Full-genome sequencing for both strains S18 and S19 presented, respectively, the following genomic constellations: G6-P[14]-I2-R2-C2-M2-A3-N2-T6-E2-H3 and G10-P[14]-I2-R2-C2-M2-A11-N2-T6-E2-H3. Phylogenetic analyses and the analysis of the VP8* antigenic epitopes for S18, S19 and ma31 revealed a shared similarity with bovine, caprine, ovine and human strains from Morocco and other countries. The VP2 and NSP1 genes of the S19 strain were closely related to those of the cognate genes of the human ma31 strain, while the VP4 gene of S18 strain was closely related to the cogent gene of the ma31 strain. Our findings revealed cases of zoonotic transmission and confirmed the risk of emergence of new genotypes in some environments such as nomadic regions, where close physical proximity between human and livestock is common. The present study is novel in reporting whole-genome analyses of RVA isolates obtained from nomadic livestock in Morocco. Electronic supplementary material The online version of this article (10.1007/s11262-020-01778-w) contains supplementary material, which is available to authorized users.

Background Several studies have shown that the genetic host factors may affect the progression of hepatitis C (HCV) and has an effect on single nucleotide polymorphisms interleukin 28B (IL28B) gene in patients with based treatment of INF/RIB. Objectives The aim of our study is to evaluate the rate of rs8099917 genotypes of IL28B and examine its impact on the responsive treatment among Moroccan patients with chronic Hepatitis C. Patients and methods A total of 173 HCV-positive people who received PEG-IFN/RBV were recruited in this study in which IL28B polymorphisms were genotyped by sequencing. Results the SVR rates of our study were 40% in patients with genotype 1 and 81% in those with genotype 2. The frequencies of the genotypes rs8099917 were 67.05% TT; 31.79% TG and 1.15% GG. Conclusion In the patients with chronic HCV-genotype 1b and 2 infections, IL28B is significantly associated with the responsive treatment of PEG-interferon/ribavirin among Moroccan patients with chronic HCV infection.

Background: Here, we describe the characterization of the VP7 and VP4 genes of rotavirus group A (RVA) strains isolated from Moroccan infants with acute gastroenteritis (AGE) after receiving the Rotarix. Methods: Stool samples were collected from seven infants suffering with AGE. Vesikari 20-point was used while collecting the AGE information. RT-PCR followed by sequencing was employed to determine partial sequences from VP7 and VP4 encoding genes. RVA genotypes were determined using the RotaC online classification tool.The Neighbor-Joining phylogenetic tree was performed in the MEGA 6.06 software as well as the amino acid (aa) alignments of VP7 and VP8* antigenic epitopes. Results: Five children had gastroenteritis episodes defined as severe on the Vesikari scale. Analysis of VP7 gene sequences revealed five G2 and two G9 genotypes. These genotypes were associated with P[4] and P[8], respectively. Phylogenetic analysis demonstrated that G2 and G9 sequences grouped together with Italian strain belonging to lineage II and III respectively. Their P[4] sequences showed only 65-73% identity and grouped into two separate sub-clusters. The Moroccan P[8] isolates did not display this correlation and segregate to lineage III. The VP8* aa of P[8] when compared with that of the Rotarix vaccine, has a six aa differences. Conclusion: Although, this study does not reveal prevalence rates of RVA in infants after the vaccine introduction, it reports the detection and genetic characteristics of the G2P[4] and G9P[8] RVA types in vaccinated children in Morocco. The findings are suggestive of continuous surveillance program on rotavirus evolution in the vaccinated population.

Reported here are the results of antigenic and genetic characterization of equine influenza strains causing local outbreaks reported in Morocco, respectively, in 1997 and 2004. The antigenic and genetic characterizations of the equine influenza virus H3N8 are reported here. The highest similarity between the HA1 nucleotide sequences of A/equine/Nador/1/1997 and those of A/equine/Rome/5/1991 and A/equine/Italy/1199/1992 demonstrate that A/equine/Nador/1/1997 belongs to the European lineage. On the other hand, A/equine/Essaouira/2/2004 and A/equine/Essaouira/3/2004 were classified in the predivergent lineage. The present work emphasizes the importance of a national influenza survey program, which requires a collaborative laboratory network to promote the collection and characterization (antigenic and genetic) of equine influenza viruses in real time.

Equine influenza is an infectious and contagious disease of horses. Studies on this topic are rare in the Maghreb countries. Therefore, the aim of this work is to present the various studies conducted on serological and molecular equine influenza virus since 1972 in the Maghreb region in particular in Morocco, Algeria and Tunisia. A total of four equine influenza strains were isolated in the Maghreb Arab region. A/equine/Nador/1/1997(H3N8), A/equine/Essaouira/2/2004(H3N8), A/equine/Essaouira/3/2004(H3N8) and A/equine/Algiers/1/1972(H3N8). The highest homology of HA nucleotide sequences of A/equine/Nador/1/1997(H3N8) with European strains: A/equine/Italy/1199/1992(H3N8) and A/equine/Brescia/1999(H3N8) clearly clustered A/equine/Nador/1/1997(H3N8) with the strains belonging to the European lineage. However, A/equine/Algiers/1/1972(H3N8), A/equine/Essaouira/2/2004(H3N8) and A/equine/Essaouira/3/2004(H3N8) were placed in the predivergent lineage indicating that like-Miami/63 strains infected equids in Morocco in 2004. This finding does not corroborate the recent studies of the H3N8 subtype of equine influenza viruses which have demonstrated that the oldest equine H3N8 strains, circulating before 1990 apparently went extinct.

Background Host genetic factors are involved in resistance/susceptibility to HIV-1 infection and progression. The most relevant genetic variant of the gene encoding HIV-1 major co- receptor, CCR5-Δ32, has a protective effect against HIV-1 infection, and the prevalence of this mutation varies by ethnicity. In Morocco, only limited information is available concerning this protective allele. Objectives The aim of this case-control study was to investigate the frequency of CCR5-Δ32 among HIV-1 infected and uninfected individuals in Morocco and to assess the association between this protective allele and HIV-1 susceptibility and disease progression. Patients and methods A total of 148 HIV-1 seropositive (cases) and 342 healthy Moroccan individuals (controls) were recruited from Mohammed V Military Training Hospital in Rabat. The CCR5-Δ32 variant was analysed by Polymerase Chain Reaction (PCR) assay. Comparison of Genotype and allele frequencies between cases and controls was performed using the χ 2 test and the Fisher exact test. Association between CCR5-Δ32 polymorphism and HIV-1 disease progression was assessed using ANOVA test. Results 7 of 148 (4.73%) HIV-1 seropositive (cases group) were heterozygous for the CCR5-Δ32 allele compared to 6 of 342 (1.75%) HIV-1 seronegative individuals (controls group). None of the analysed individuals had CCR5-Δ32 homozygous genotype. No statistical significant association was found in the genotype distribution between cases and controls. Also, we didn't observe any significant association between CCR5-Δ32 polymorphism and the markers of HIV-1 disease progression (HIV-1 viral load or CD4 cell counts) (p > 0.05). Conclusion The present results are the first findings on the association of CCR5-Δ32 variant allele with HIV-1 susceptibility and the markers of HIV-1 disease progression among Moroccan HIV-1 seropositive patients. These results suggest that CCR5-Δ32 mutation may not affect HIV-1 susceptibility in heterozygous individuals. Given the low frequency of the Δ32 mutated allele in our population, further studies are needed to confirm our results and evaluate the relationship between CCR5-Δ32 polymorphism and HIV-1 disease progression in Morocco.

Background West Nile virus (WNV) emerged in Morocco in 1996 with a re-occurrence in 2003, 2010 and recently in 2015 in specific areas through the country. Here, we conducted a serosurvey on Moroccan military individuals to evaluate blood donors at risk for having WNV. Methods During a period of West Nile Virus (WNV) activity (from …

During the last fifteen years, new viral diseases such Bluetongue (BT), West Nile (WN), African horse sickness (AHS), Epizootic hemorrhagic disease (EHD) and Peste des Petits Ruminants (PPR) have extended their geographic distribution and emerged in North Africa and in Europe. Camel (Camelus dromedarius) is considered as a potential vector in the transmission of some of these diseases while it is host-specific for Camelpox (CP). A serological survey was conducted on 1392 sera to estimate changes of these diseases prevalence in camels over two different time spans (2003 and 2009) and across different sites of South Morocco. Results indicate clearly that BT was circulating in camels before the first notified outbreak in Morocco (2004) with 42% positive sera in Guelmim in 2003. BT and WN prevalence's increased over time from 11 to 25% and from 10 to 13% respectively. Higher prevalence of both diseases was found in camels living in oases indicating an increased intensity of viral circulation in these sites. Camels have been tested negative for AHS, EHD and PPR viruses while higher CP prevalence has been detected in camels living in Smara, the most closed site to eastern borders (i.e. Mauritania). The seroprevalence of CP in camels at interval of 7 years increases from 23% to 37%. This survey could provide information on the possible use of camel as a sentinel animal for surveillance of emerging diseases such BT and WN.

Summary The potential role of domestic dogs in the long-distance transmission of bluetongue virus (BTV) is currently unproven. This study set out, through an experimental infection study, to investigate whether domestic dogs mount a viraemia post-infection with a field strain of BTV serotype 1. All six experimentally infected dogs seroconverted within 14 days and viral RNA was detected in the blood of the dogs, albeit at significantly lower levels than that seen in domestic ruminants. There was no clear evidence for viral replication in the dogs as no increase in viral RNA was observed in, and it was not possible isolate virus from, the blood of the dogs. There was however evidence for a persistence of viral RNA in the blood of the dogs, which may be evidence for a low level of replication or could be indicative of persistence of the viral inoculum.

Dengue is a significant mosquito-borne infection in humans, and its worldwide prevalence is rapidly increasing. The vector aedes albopictus has been revealed recently in the town of Rabat. Morocco established a program of active surveillance of dengue fever comprising many hospitals and laboratories across the kingdom. The purpose of this work is to describe the result of the surveillance of the dengue virus (DENV) infection during the year of 2017 among Moroccans and tourists who presented in our hospital with clinical signs of infection and to report the first confirmed positive cases of Dengue.From 20 December 2016 to 20 December 2017, 21 patients were hospitalized for suspicion of DENV infection. Half of them were returning from Côte d’Ivoire which is a popular tourist and business country for Moroccans and where an outbreak of DENV was confirmed on July 2017. Fever, headache, arthralgia-myalgia and malaise in addition to the notion of return from an endemic country justify in clinicians the demand for analysis of detection of dengue virus by RT-PCR.Dengue infection was confirmed in two patient both coming from Côte d’Ivoire, a Moroccan and an Ivorian who were staying in Abidjan during the period of the outbreak of 2017.

Recently, several countries reported imported cases of infection with chikungunya virus (CHIKV). We report the first case of chikungunya virus infection in Morocco. A 37-year old woman returned to Morocco on 15 August 2017, after she stayed in Dhaka-Bangladesh for 18months. She developed severe arthralgias and rash, fever up to 39°c. In next day’s symptoms progressively subsided but arthralgias remained for 3weeks. Laboratory findings didn't show lymphopenia, thrombocytopenia or elevated liver transaminases. Serological tests were positive for CHIKV IgM and negative for IgG antibodies. CHIKV-RNA was detected by RT-PCR. The patient was treated with non-steroid anti-inflammatory drugs and paracetamol. After 15days of hospitalization, symptoms ameliorated but arthralgias persists. The vector is established in Morocco and since the virus is diagnosed in returning travellers, chikungunya has a potential for autochthonous transmission in Morocco, that’s why CHIKV must be included in the differential diagnosis of arthralgia in all travellers returning from countries with documented transmission of the virus.

Background The equine influenza (EI) is an infectious and contagious disease of the upper respiratory tract of horses. Two outbreaks were notified in Morocco during 1997 and 2004 respectively in Nador and Essaouira. The aims of the present study concern the amino acids sequences comparison with reference strain A/equine/Miami/1963(H3N8) of the HA2 subunit including the cleavage site of three equine influenza viruses (H3N8) isolated in Morocco: A/equine/Nador/1/1997(H3N8), A/equine/Essaouira/2/2004 (H3N8) and A/equine/Essaouira/3/2004 (H3N8). Results The obtained results demonstrated that the substitutions were located at Ectodomain (ED) and transmembrane domain (TD), and they have only one arginine in cleavage site (HA1-PEKQI-R329-GI-HA2). In the Ectodomain, the mutation N/154 2 /T deleted the NGT glycosylation site at position 154 for both strains A/equine/Essaouira/2/2004(H3N8) and A/equine/Essaouira/3/2004(H3N8). Except for mutation D/1602/Y of the A/equine/Nador/1/1997(H3N8) strain, the other mutations were involved in non conserved sites. While the transmembrane domain (TM) of the strain A/equine/Essaouira/3/2004(H3N8) exhibits a substitution at residue C/199 2 /F. For the A/equine/Nador/1/1997(H3N8) strain the HA2 shows a mutation at residue M/207 2 /L. Three Moroccan strains reveals a common substitution at the residue E/211 2 /Q located between transmembrane domain TM and the cytoplasmic domain (CD). Conclusion The given nature virulence of three Moroccan strains, the identified and reported mutations certainly played a permissive role of infection viral process.

Moviegoers commonly use film critics as information sources. This article proposes that consumers engage in a causal analysis of the reasons behind critics' judgments and recommendations about new movies before making their own evaluations. An experimental study was conducted where critic's style, critic's predisposition toward the film director, degree of agreement between the critic's judgment and that of other critics, reputation of the critic, reputation of the film director, and critic's judgment direction were manipulated in a mixed within-subjects factorial design. A summary movie review was constructed for each condition of the factorial design. Viewer's movie evaluations were assessed with the use of four evaluative bipolar scales. Three of the five research hypotheses were fully supported. Consistent with the predictions of attribution theory, it was found that consumers' evaluations of a new movie are more likely to follow the judgment of a film critic when this judgment does not conform to the critic's style (H1), when the judgment is inconsistent with the critic's predisposition toward the film director (H2) and when other critics' judgments show favorable consensus (H3). The other two research hypotheses were partially supported. The impact of the film director's reputation on consumer evaluations (H5) was significant only when the movie reviews were positive, whereas the impact of the critic's reputation (H4) was significant only in the case of negative reviews. The results are interpreted within the framework of attribution theory. © 1999 John Wiley & Sons, Inc.

Experimental challenge protocols that consistently reproduce clinical signs of peste des petits ruminants in Alpine goats infected with a tissue culture-passaged peste des petits ruminants virus are described. The protocols can be used to carry out quality-controlled vaccine efficacy and pathogenesis studies under experimental conditions.

Peste des petits ruminants virus (PPRV) recently caused a serious outbreak of disease in Moroccan sheep and goats. Alpine goats were highly susceptible to PPRV with mortality rates approaching 100%, as opposed to local breeds of sheep which were less susceptible to the disease. The relative susceptibility of alpine goats was investigated through an experimental infection study with the Moroccan strain of PPRV. Severe clinical signs were observed in the alpine goats with virus being excreted through ocular, nasal and oral routes. No difference in the severity of the disease in goats was observed with different inoculation routes and transmission of the virus by direct contact was confirmed. This study confirmed the susceptibility of the alpine goat to PPRV infection and describes a challenge protocol that effectively and consistently reproduced severe clinical signs of PPR in experimentally infected goats.

The aim of the present study was to observe the induction and repair of single strand breaks (Ssbs) and double strand breaks (Dsbs) in mesophyll protoplasts of Nicotiana plumbaginifolia, irradiated with UV-C and cultured under light or dark conditions. DNA damage and repair was determined by the neutral and alkaline comet assay to reveal Dsbs and Ssbs respectively.

The first objective of our study was to analyse whether biomarkers for genotoxic effects (DNA breaks and alkali-labile sites and micronucleus and non-disjunction frequencies) could be fully validated for biomonitoring workers chronically exposed to ionizing radiation (IR). Blood samples of controls and individuals chronically exposed to IR were analysed. The interindividual variation was reduced when the comet data were adjusted for interexperimental variation, but remained statistically significant. No differences were found between groups, either for smoking or for exposure status. The second objective was to determine whether the Comet assay can be used to evaluate global repair phenotype as a susceptibility biomarker for IR-induced DNA damage in nuclear workers. A pilot study was performed and blood from workers exposed or not to radiation was submitted to a challenging dose of gamma-rays. The repair kinetics of each individual donor were analysed by Comet assay at different time points and compared with the frequencies of biomarkers of genotoxic effects. There was a statistically significant interaction between biomarkers assessing the same damage (micronucleus and Comet assays). Multivariate analysis showed that micronucleus frequencies were positively influenced by age and the percentage of residual tail length was negatively influenced by the interaction between smoking and exposure status. The general conclusions from our study are: (i) a positive correlation exists between mechanistically related biomarkers; (ii) multivariate regression analysis confirmed that the interaction between smoking and exposure to IR negatively and statistically significantly influenced residual tail length; (iii) use of the Comet assay for the estimation of global repair phenotype with respect to IR is recommended because it is simple, fast and differences in in vitro repair capacity can be detected.

The outcome of liver transplant recipients in HCV chronic carriers with Anti-HBc only concerning occult HBV infection is unknown. We report here the case of a patient who underwent liver transplantation (LT) for cirrhosis post chronic hepatitis C who received an allograft from a donor with no marker of hepatitis B infection. After LT, HBV DNA was detected in the serum in the absence of HBsAg while HCV RNA remained negative. To determine the origin of this occult HBV infection, we retrospectively examined stored serum and liver tissue, pre and post-transplantation, for HBV DNA by PCR. A stored liver biopsy of the donor before transplantation was also tested. HBV DNA was detected in the pre-transplant liver but not in the donor liver. HBV viral load quantified by real time PCR after LT ranged from about 10 2 to 5x10 3 HBV DNA copies/mg of liver, while in sera, concentrations ranged from 10 2 to 3x10 3 HBV DNA copies/ml. All PCR products in the S gene from liver and sera were sequenced. Analysis of sequences showed the presence of an HBV strain genotype D. The nucleotide homology between the patient’s HBV strains before and after LT was 96 % across the analyzed regions. Full length HBV genomes were amplified from the sera using Rolling Circle Amplification and then sequenced. Analysis of sequences confirmed the genotype D, but did not show obvious mutations that could contribute to HBsAg seronegativity and low HBV viral replication. Factors leading to occult HBV infection are still unclear, but it is well establish that occult HBV infection is frequent in HCV patients. This underlines the role of extra hepatic sites for HBV replication, potentially lymphocytes acting as “reservoirs”.

The comet assay is widely used to detect DNA damage in single cells. However, only moderate attention has been paid to the experimental variability of this assay, especially during electrophoresis. To take into account this variation and to be able to compare measurements from different electrophoretic runs, as would be necessary when large numbers of samples need to be analysed, it is important to integrate an internal standard into the assay. This study presents a first step in the validation and implementation of an internal standard in the alkaline comet assay. Untreated and ethyl methanesulfonate treated cells (K562 human erythroleukemia cell line) were used as negative and positive internal standards, respectively, in each electrophoresis run. Three steps were followed: (1) assessment of the different levels of variability which may influence the damage levels of the internal standards, (2) evaluation of the variability across separate electrophoresis runs on the quantification of DNA damage in the internal standards by three experimenters involved in different studies and (3) proposal of an adequate calculation system to integrate the internal standards into test sample data. The application of the two proposed models to samples from a human biomonitoring study is presented. The model which calibrates the measurements against the negative internal standard is the most useful since this negative standard was the most stable across experiments and among the three experimenters. The percentage of DNA in the tail is the most appropriate parameter to analyse induced DNA damage, because its interelectrophoresis and interexperimenter variation is less pronounced than that of tail length.

The evaluation of different cytogenetic endpoints of radiation damage for the biomonitoring of contract workers temporarily employed at nuclear power plants.Blood samples from six donors were irradiated in vitro with doses ranging from 0.1 to 2Gy 60Co gamma-rays. Compared were a conventional analysis for dicentrics, the conventional micronucleus (MN) assay, the centromere micronucleus assay using p82H and an alphaAllCen pancentromeric probe, and tricolour FISH with chromosome 2, 4 and 8 DNA probes for the scoring of translocations.Agreement in the number of MN between Giemsa-and propidium iodine fluorescence-stained preparations was obtained. The control samples showed higher centromere positivity for the MN after FISH with the p82H probe compared with the alphaAllCen probe. The MN results with both probes showed a slight but systematic increase in the number of centromere-positive MN with dose, indicating that radiation, although principally clastogenic, also has aneuploidogenic properties. The values of the genomic translocation frequency (FG) derived from the observed translocation frequencies were systematically higher than the dicentric yields. Comparing the sensitivity of the different methods with restriction of the scoring time to 1 day for biomonitoring purposes, the centromere micronucleus assay had the lowest dose detection limit (0.1 to 0.2 Gy).This study shows that at present only the centromere micronucleus assay can combine high sensitivity with a reasonable scoring time for the biomonitoring of relatively large populations.