A short artificial antimicrobial peptide shows potential to prevent or treat bone infections.

Infection of bone is a severe complication due to the variety of bacteria causing it, their resistance against classical antibiotics, the formation of a biofilm and the difficulty to eradicate it. Antimicrobial peptides (AMPs) are naturally occurring peptides and promising candidates for treatment of joint infections. This study aimed to analyze the effect of short artificial peptides derived from an optimized library regarding (1) antimicrobial effect on different bacterial species, (2) efficacy on biofilms, and (3) effect on osteoblast­like cells. Culturing the AMP-modifications with Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus aureus (including clinical isolates of MRSA and MSSA) and Staphylococcus epidermidis identified one candidate that was most effective against all bacteria. This AMP was also able to reduce biofilm as demonstrated by FISH and microcalorimetry. Osteoblast viability and differentiation were not negatively affected by the AMP. A cation concentration comparable to that physiologically occurring in blood had almost no negative effect on AMP activity and even with 10% serum bacterial growth was inhibited. Bacteria internalized into osteoblasts were reduced by the AMP. Taken together the results demonstrate a high antimicrobial activity of the AMP even against bacteria incorporated in a biofilm or internalized into cells without harming human osteoblasts.

Complete Nucleotide Sequence of Artichoke latent virus Shows it to be a Member of the Genus Macluravirus in the Family Potyviridae.

Complete genomic sequences of Artichoke latent virus (ArLV) have been obtained by classical or high-throughput sequencing for an ArLV isolate from Italy (ITBr05) and for two isolates from France (FR37 and FR50). The genome is 8,278 to 8,291 nucleotides long and has a genomic organization comparable with that of Chinese yam necrotic mosaic virus (CYNMV), the only macluravirus fully sequenced to date. The cleavage sites of the viral polyprotein have been tentatively identified by comparison with CYNMV, confirming that macluraviruses are characterized by the absence of a P1 protein, a shorter and N-terminally truncated coat protein (CP). Sequence comparisons firmly place ArLV within the genus Macluravirus, and confirm previous results suggesting that Ranunculus latent virus (RALV), a previously described Macluravirus sp., is very closely related to ArLV. Serological relationships and comparisons of the CP gene and of the partial RaLV sequence available all indicate that RaLV should not be considered as a distinct species but as a strain of ArLV. The results obtained also suggest that the spectrum of currently used ArLV-specific molecular hybridization or polymerase chain reaction detection assays should be improved to cover all isolates and strains in the ArLV species.

First Report of Tomato torrado virus in Tomato Crops in France.

In June 2008, tomato (Solanum lycopersicum L.) plants cv. Fer De Lance (De Ruiter Seeds, Bergschenhoek, the Netherlands) grown in greenhouses near Perpignan (southern France) showed growth reduction and necrotic lesions on fruits, stems, and basal parts of the leaves. Tomato torrado virus (ToTV) was suspected on the basis of symptoms and its recent description in Spain (4). Primer set A (3), designed to ToTV RNA-2, was used for reverse transcription (RT)-PCR experiments on RNA extracted from four infected plants and allowed the amplification of a 493-bp fragment. No amplification was observed from healthy plant extracts. The RT-PCR product was directly sequenced (GQ303330) and a BLAST search in GenBank revealed 99.8- and 99.5%-nt identity with Polish (EU563947) and Spanish type strain (DQ388880) isolates of ToTV, respectively. Double-antibody sandwich-ELISA tests were conducted on these four samples to check for the presence of other viruses commonly found in tomato crops in France. Tomato spotted wilt virus, Parietaria mottle virus, Cucumber mosaic virus, Tomato mosaic virus, and Potato virus Y were not detected but Pepino mosaic virus (PepMV) was detected in all samples. ToTV was mechanically transmitted to Physalis floridana but PepMV was not. This plant was used to inoculate healthy tomatoes that served as a ToTV source for further experiments. Mechanical inoculation to test plants showed that Nicotiana benthamiana, N. clevelandii, N. debneyi, N. glutinosa, Capsicum annuum, Solanum melongena, and some tomato cultivars (including Fer De Lance), in which typical necrotic symptoms were observed, were systemically infected by the virus. Isometric particles ~28 nm in diameter were observed by electron microscopy in crude extracts of infected plants negatively stained with 1% ammonium molybdate, pH 7. To confirm ToTV identification, whitefly transmission experiments were performed with Trialeurodes vaporariorum and Bemisia tabaci. Adult whiteflies were placed in cages with infected tomato plants for 1-, 24-, or 48-h acquisition access periods (AAP) before transferring them by groups of ~50 on susceptible tomato plantlets placed under small containers (six plants per AAP). Forty-eight hours later, plants were treated with an insecticide and transferred to an insect-proof containment growth room. Ten days later, RNA preparation from all plants was tested by RT-PCR for the presence of ToTV. No transmission was observed with a 1-h AAP. With a 24-h AAP, transmission to four of six test plants was observed with both whitefly species, while at 48 h, AAP transmission to three and four plants of six was observed with T. vaporariorum and B. tabaci, respectively. Noninoculated control plants were all negative by RT-PCR. These experiments confirm T. vaporariorum and B. tabaci as natural vectors of ToTV as previously described (1,2). ToTV has been already reported in Spain, Poland, Hungary, and Australia, but to our knowledge, this is the first report of ToTV in France. Our detection of ToTV in April 2009 from the same area revealed 7 positive tomato plants of 17 tested. This observation suggests the persistence of the disease in the Perpignan Region. References: (1) K. Amari et al. Plant Dis. 92:1139, 2008. (2) H. Pospieszny et al. Plant Dis. 91:1364, 2007 (3) J. Van der Heuvel et al. Plant Virus Designated Tomato Torrado Virus. Online publication. World Intellectual Property Organization WO/2006/085749, 2006. (4) M. Verbeek et al. Arch. Virol. 152:881, 2007.

First Report of Cucurbit yellow stunting disorder virus in Commercial Cucumber Greenhouses in France.

In autumn 2001, severe yellowing symptoms were observed on greenhouse-grown cucumbers near Perpignan (southern France). Leaf samples were collected from two sites where plants displayed symptoms ranging from limited yellowing of the older leaves to severe, complete yellowing of the whole plant. Cucurbit aphid-borne yellows virus, a polerovirus that causes similar symptoms was not detected in doubleantibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using a specific antiserum. Total RNA was extracted from fresh leaf tissues and used in reverse transcription-polymerase chain reaction (1) with primers specific for two whitefly-borne viruses also inducing yellows and occurring in the Mediterranean basin (1): Beet pseudo yellows virus (BPYV, genus Closterovirus) transmitted by Trialeurodes vaporariorum (West.) and Cucurbit yellow stunting disorder virus (CYSDV, genus Crinivirus) transmitted by Bemisia tabaci (Genn.). No BPYV was detected in this survey, but CYSDV was present in all samples. In subsequent surveys conducted in the spring and summer of 2002, BPYV and CYSDV were detected, sometimes in mixed infections, in samples collected from the same region. The complete CYSDV coat protein gene was amplified by PCR using specific primers (2), yielding the expected-size fragment of 756 bp. The French isolate (GenBank Accession No. AY204220) shared 99.6 to 100% nucleotide sequence identity in the sequenced CP fragments (700 nt) with isolates of the most common, highly homogenous subgroup of CYSDV that has emerged recently in the Middle East, southwestern Europe (Spain and Portugal), United States, and Morocco (2). To our knowledge, this is the first report of CYSDV in France and it shows the threat represented by the current emergence of B. tabaci-transmitted viruses. References: (1) I. C. Livieratos et al. Plant Pathol. 47:362, 1998. (2) L. Rubio et al. J. Gen. Virol. 82:929, 2001.

Obstructive sleep apnea: a comparison of continuous positive airway pressure and surgical treatment.

Since earlier descriptions of the syndrome of obstructive sleep apnea (OSA), various treatment alternatives have included a variety of medical regimens, continuous positive airway pressure (CPAP), tracheostomy, and other surgical options. A lack of acceptable criteria for surgical intervention remains an important concern for the surgeon. in an attempt to resolve some of the controversies pertaining to various therapeutic modalities, we performed a retrospective analysis--from 1983 to the present--of posttreatment results in patients who underwent surgical therapy and those who were treated primarily with CPAP at this institution. Of 400 patients diagnosed with OSA, only 66 underwent surgical treatment, including uvulopalatopharyngoplasty. CPAP was the mainstay of treatment in the majority of our patients. Post-treatment data were available for 50 patients treated with CPAP and for 45 patients treated surgically. A comparative analysis of polysomnographic studies revealed superior cures with CPAP, although long-term compliance remains a significant problem. We advocate CPAP as initial therapy in patients with no clinically apparent causes for obstruction (e.g., nasal polyps, deviated nasal septum, or obstructive tonsillar hypertrophy) because of the predictability of success, and lower costs and complication rates. Long-term followup of OSA patients is indicated, regardless of treatment modality.

Upper-airway surgery for treating obstructive sleep apnea. Measuring and predicting success.

The effectiveness of upper-airway surgery in treating obstructive sleep apnea was examined in 37 patients. All patients received a uvulopalatopharyngoplasty. Some also received tonsillectomies, septoplasties, and turbinectomies. The success of the operations was evaluated along three dimensions: a decrease in the number of apneas per sleep hour, a reduction in maximum nocturnal oxygen desaturation, and a decrease in daytime somnolence. Over 80% of the patients improved on at least one of these factors following surgery. However, postoperative intercorrelations among these three variables were relatively low. Furthermore, preoperative indexes of apnea severity could not be used to predict the degree of postoperative improvement.

Nightmares in Vietnam veterans.

Repetitive traumatic dreams remain a curiosity in our working with patients. If dreams have a sleep protecting or problem solving function, the dream that does not resolve but repeats and that disturbs rather than protects sleep remains an important clinical problem. We see in individual patient experiences and in civilian disasters samples of individuals who are unable to return their dream life to a condition in which it no longer repeats or disturbs their sleep. Clearly the recent experience of combat survivors of Vietnam becomes a major concern.

Choosing between predictable shock schedules: Long- versus short-duration signals.

Two experiments assessed the relative aversiveness of different duration preshock signals (5 and 20 seconds) and different duration stimuli identifying shock-free periods. In the first experiment, the responding of 15 of 18 rats was maintained when it produced changes from a predictable-shock condition with a 5-second preshock signal to an identical schedule with a 20-second preshock signal; responding was not maintained when it produced the opposite changes. These results occurred with intershock intervals of both 120 seconds and 240 seconds. The second experiment assessed whether changing to the 20-second schedule was maintained by properties of the preshock signals identifying the shock periods or by properties of the stimuli identifying the shock-free periods. Four subjects were given training with the two signaled schedules in an operant chamber and then later given off-baseline preference tests in a shuttlebox. When given a choice between preshock signals, subjects chose the 5-second signals over the 20-second signals. However, when given a choice between stimuli identifying shock-free periods, subjects chose the stimulus identifying the shorter shock-free periods (i.e., the one previously correlated with the 20-second signals). These findings are discussed within the Rescorla and Wagner model of stimulus compounds and within the context of safety as a contrast phenomenon.

Problems in the use of long-acting hypnotics in older patients.

Insomnia and the use of long-acting hypnotic drugs are prevalent in geriatric populations. These drugs have been shown in the laboratory to adversely affect motor performance in the elderly. To examine the clinical significance of these findings, comparisons were made between patients who experienced a fall during their hospital stay and those who did not fall. It was found that patients over 70 years of age who experienced a fall in the hospital were more likely to be receiving flurazepam than the patients who did not fall.