IAPV, a bee-affecting virus associated with Colony Collapse Disorder can be silenced by dsRNA ingestion.

Colony Collapse Disorder (CCD) has been associated with Israeli acute paralysis virus (IAPV). CCD poses a serious threat to apiculture and agriculture as a whole, due to the consequent inability to provide the necessary amount of bees for pollination of critical crops. Here we report on RNAi-silencing of IAPV infection by feeding bees with double-stranded RNA, as an efficient and feasible way of controlling this viral disease. The association of CCD with IAPV is discussed, as well as the potential of controlling CCD.

Mutations in human DNA polymerase eta motif II alter bypass of DNA lesions.

Human DNA polymerase eta (hPol eta) is one of the newly identified Y-family of DNA polymerases. These polymerases synthesize past template lesions that are postulated to block replication fork progression. hPol eta accurately bypasses UV-associated cis-syn cyclobutane thymine dimers in vitro and contributes to normal resistance to sunlight-induced skin cancer. We describe here mutational analysis of motif II, a highly conserved sequence, recently reported to reside in the fingers domain and to form part of the active site in Y-family DNA polymerases. We used a yeast-based complementation system to isolate biologically active mutants created by random sequence mutagenesis, synthesized the mutant proteins in vitro and assessed their ability to bypass thymine dimers. The mutability of motif II in 210 active mutants has parallels with natural evolution and identifies Tyr52 and Ala54 as prime candidates for involvement in catalytic activity or bypass. We describe the ability of hPol eta S62G, a mutant polymerase with enhanced activity, to bypass five other site-specific lesions. Our results may serve as a prototype for studying other members of the Y-family DNA polymerases.

Transcription factor BETA2 acts cooperatively with E2A and PDX1 to activate the insulin gene promoter.

The insulin gene is efficiently expressed only in pancreatic beta cells. Using reverse transcriptase-polymerase chain reaction analysis, we show that insulin mRNA levels are at least 10(5)-fold higher in beta cells than non-beta cells. To examine the underlying mechanisms, we expressed beta cell transcription factors by transfection of non-beta cells. Separate expression of BETA2, E2A, or PDX1 led to modest (<10-fold) activation of the insulin promoter, whereas co-expression of the three proteins produced synergistic, high level activation (160-fold). This level of activity is approximately 25% that observed in transfected beta cell lines. Of the three factors studied, BETA2 appears to play a dominant role. Efficient transcription required a C-terminal activation domain of BETA2 and an N-terminal region, which does not function as an independent activation domain. The myogenic basic helix-loop-helix (bHLH) protein MyoD was unable to bind and activate the promoter, even when its DNA binding region was replaced with that of BETA2. Our results demonstrate the central importance of BETA2 in insulin gene transcription and the importance of sequences outside the canonical DNA binding domain in permitting efficient DNA binding and cell-specific activity of the insulin gene promoter.

Detection of adeno-associated virus type 2 sequences in the human genital tract.

Adeno-associated virus (AAV) is a defective parvovirus with unknown pathogenicity. It requires helper functions for its normal replication in human tissue and therefore is not readily isolated from clinical specimens. We have used the PCR method to examine the following clinical samples for the presence of AAV sequences: (i) 15 nasopharyngeal aspirates from symptomatic patients, (ii) 7 swab or fluid specimens from vesicles of patients suspected of having varicella-zoster virus infections, (iii) 21 human papilloma virus-positive genital biopsy specimens, (iv) 61 genital swab specimens from women suspected of having herpes simplex virus (HSV) infection examined either directly or following propagation in tissue culture, (v) 62 samples of first-trimester aborted material, including 38 samples from spontaneous abortions and 24 samples from induced abortions, (vi) 11 samples of chorionic villi taken from women undergoing genetic prenatal diagnosis, and (vii) three lots of cultured human embryonic cells. AAV sequences were detected only in samples taken from the genital tracts of women suspected of having HSV infection and not in any of the other types of samples. Samples from 11 patients were positive for AAV: for 4 patients the original swab sample was positive, for 4 patients the cultured swab sample was positive, and for 3 patients both the original swab samples and the cultures were positive. Five of the 11 patients were infected with HSV. Our study demonstrates the presence of AAV in the female genital tract. However, in contrast to a previous report (E. Tobiasch, M. Rabreau, K. Geletneky, S. Larue-Charlus, F. Severin, N. Becker, and J. R. Schlehofer, J. Med. Virol. 44:215-222, 1994), we did not find solid evidence of its replication in maternal or embryonal tissues from the first trimester of pregnancy. The questions of a potential pathogenic etiology of AAV and the interaction with HSV remain open.

Agrobacterium radiobacter peritonitis in two patients maintained on chronic peritoneal dialysis.

We report two patients with end-stage renal disease maintained on chronic peritoneal dialysis who developed peritonitis in which the infecting organism was Agrobacterium radiobacter, normally a rare pathogen in humans. Both patients initially responded to antibiotics, but later relapsed and required catheter removal. Neither had been exposed to soil or plant material. A radiobacter is yet another of a growing list of unusual organisms that infect the peritoneal cavity of peritoneal dialysis patients.

In vitro activity of tosufloxacin against bacterial enteric pathogens.

The in vitro activity of tosufloxacin (A-61827), a new quinolone antibiotic, was compared with that of four other quinolones against 162 bacterial enteric pathogens. Susceptibility testing was performed by using an agar dilution method. The minimal inhibitory concentration (MIC) was defined as the lowest concentration of antibiotic without visible growth. Tosufloxacin was the most active agent against Campylobacter jejuni and C. coli, and ciprofloxacin was the most active agent against Salmonella, Shigella, and Vibrio. The frequency of spontaneous point mutational resistance to tosufloxacin for three C. jejuni ranged from 1.2 x 10(-9) to 5.1 x 10(-13). No significant differences in mutational frequency were seen among the quinolones.

Reflex sympathetic dystrophy (algoneurodystrophy): temperature studies in the upper limb.

The temperature response of the hands to mild cold stress (20 degrees C for one minute) has been measured in 20 normal subjects, 20 patients with reflex sympathetic dystrophy (RSD) and 10 patients with chronic upper limb pain (CULP) of uncertain origin. The results of RSD and CULP groups were significantly (p less than 0.05) different from normal but were indistinguishable. For each patient, 11 variables obtained from the thermal stress test were compared with the normal range. Ten of the RSD group and seven of the CULP group had four or more abnormal variables and were considered to have a thermoregulatory abnormality. The thermal stress test is useful in the objective assessment of RSD. It is non-invasive, patient acceptable and reproducible.

A multi-centre study of sulindac versus naproxen in the treatment of elderly osteoarthritic patients.

Ninety-five geriatric patients with osteoarthritis of the hip and/or knee were entered into an open parallel group study of sulindac (400 mg/day) and naproxen (500 mg/day), both given in twice-daily regimens. Patients received the drugs for 12 weeks. Both drugs produced improvements in the patients' overall condition. There were no statistically significant differences between the effects of the two drugs. Overall, both drugs proved beneficial and well tolerated.

Post-traumatic neurodystrophy. Treatment by corticosteroids.

This paper reports the results with corticosteroid therapy given to patients with neurodystrophy who failed to improve on other treatment, including intensive physiotherapy.

A clinical Trial of Tofranil in osteo-arthritis.

Animal experiments and uncontrolled clinical experience indicate that imipramine should relieve pain or enhance the effect of analgesics in osteoarthritis. A double-blind crossover trial using Tofranil 25 mg three times daily and matching placebo has, therefore, been designed and initiated.

Myasthenia gravis associated with penicillamine treatment for rheumatoid arthritis.

Four patients with rheumatoid arthritis (R.A.) developed myasthenia gravis after taking penicillamine. In one patient withdrawal of the drug was followed by spontaneous remission of the myasthenia, and in two the dose of anticholinesterase was subsequently reduced. In the fourth patient continuing penicillamine treatment was associated with increasingly severe myasthenic features, but on withdrawal of the drug these resolved. As myasthenia gravis rarely complicates R.A. its onset in these patients shortly after the start of penicillamine treatment suggested that penicillamine may have precipitated this condition.