Challenges and opportunities in evaluating a digital sexual health intervention in a clinic setting: Staff and patient views.

OBJECTIVES: The aim of the study was to gather the views of sexual health clinic staff and male clinic users regarding digital sexual health promotion and online trial procedures. METHODS: The Men's Safer Sex website was offered on tablet computers to men in the waiting rooms of three sexual health clinics, in a feasibility online randomised controlled trial (RCT). Interviews were conducted with 11 men who had participated in the trial and with nine clinic staff, to explore their views of the website and views of the online trial. Interviews were audio-recorded and transcribed, and we conducted a thematic analysis of interviews and of 281 free text comments from the online RCT outcome questionnaires. RESULTS: Clinic users and staff felt that digital interventions such as the Men's Safer Sex website are useful, especially if NHS endorsed. Pre-appointment waiting time presents a good opportunity for intervention but clinic users and staff felt that a website should supplement rather than replace face-to-face healthcare. The RCT procedures fitted well around clinical activities, but men did not self-direct to the tablet computers. Staff were more concerned about consent and confidentiality than clinic users, and staff and patients were frustrated by multiple technical problems. The trial outcome questionnaire was thought-provoking and could constitute an intervention in itself. Participants felt that clinics would need to promote a digital intervention and/or offer the site routinely to promote engagement. CONCLUSION: Digital interventions could usefully supplement in-person sexual health care, but there are important obstacles in terms of IT access in NHS settings, and in promoting engagement.

Fructose content and composition of commercial HFCS-sweetened carbonated beverages.

OBJECTIVE: The obesigenic and related health effects of caloric sweeteners are subjects of much current research. Consumers can properly adjust their diets to conform to nutritional recommendations only if the sugars composition of foods and beverages is accurately measured and reported, a matter of recent concern. We tested the hypothesis that high-fructose corn syrup (HFCS) used in commercial carbonated beverages conforms to commonly assumed fructose percentages and industry technical specifications, and fulfills beverage product label regulations and Food Chemicals Codex-stipulated standards. DESIGN: A high-pressure liquid chromatography method was developed and verified for analysis of sugars in carbonated beverages sweetened with HFCS-55. The method was used to measure percent fructose in three carbonated beverage categories. Method verification was demonstrated by acceptable linearity (R(2)>0.99), accuracy (94-104% recovery) and precision (RSD < 2%). RESULT: Fructose comprised 55.58% of total sugars (95% confidence interval 55.51-55.65%), based on 160 total measurements by 2 independent laboratories of 80 randomly selected carbonated beverages sweetened with HFCS-55. The difference in fructose measurements between laboratories was significant but small (0.1%), and lacked relevance. Differences in fructose by product category or by product age were not statistically significant. Total sugars content of carbonated beverages showed close agreement within product categories (95% confidence interval = 0.01-0.54%). CONCLUSIONS: Using verified analytical methodology for HFCS-sweetened carbonated beverages, this study confirmed the hypothesis that fructose as a percentage of total sugars is in close agreement with published specifications in industry technical data sheets, published literature values and governmental standards and requirements. Furthermore, total sugars content of commercial beverages is consistent with common industry practices for canned and bottled products and met the US Federal requirements for nutritional labeling and nutrient claims. Prior concerns about composition were likely owing to use of improper and unverified methodology.

Fate of bacterial pathogens and indicator organisms in liquid sweeteners.

The survival of pathogenic and indicator microorganisms in liquid sweeteners was studied. Seven sweeteners--liquid sucrose, 42% high-fructose corn syrup (HFCS), 55% HFCS, 25 DE (dextrose equivalent) corn syrup (CS), 36 DE CS, 63 DE CS, 50% medium invert sucrose, and 65% high-maltose corn syrup (HMCS) were inoculated with Salmonella spp., Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and coliforms at a level of 10(5) cells per g. The inoculated products were stored both at or near their normal holding temperatures (32 to 46 degrees C) and at 26.7 degrees C (the lower limit during transportation). In most of the products the number of microorganisms fell below the detection limit in less than 3 days when the sweeteners were stored at their normal holding temperatures. However, in liquid sucrose S aureus survived up to 2 weeks. When the products were stored at 26.7 degrees C, the reduction in the number of microorganisms occurred at a slower rate. At 26.7 degrees C the fastest rates of reduction were observed in 42 and 55% HFCS and in 50% medium invert sucrose. In these products the number of bacteria fell below the detection limit in 3 to 6 days. The slowest rate of the reduction was observed in the liquid sucrose, in which S. aureus survived up to 1 month. These results indicate that incidental contamination of liquid sweeteners with microbial pathogens will not present a public health or regulatory hazard.

The 5'-exonuclease activity of bacteriophage T4 RNase H is stimulated by the T4 gene 32 single-stranded DNA-binding protein, but its flap endonuclease is inhibited.

Bacteriophage T4 RNase H is a 5'- to 3'-nuclease that has exonuclease activity on RNA.DNA and DNA.DNA duplexes and can remove the pentamer RNA primers made by the T4 primase-helicase (Hollingsworth, H. C., and Nossal, N. G. (1991) J. Biol. Chem. 266, 1888-1897; Hobbs, L. J., and Nossal, N. G. (1996) J. Bacteriol. 178, 6772-6777). Here we show that this exonuclease degrades duplex DNA nonprocessively, releasing a single oligonucleotide (nucleotides 1-4) with each interaction with the substrate. Degradation continues nonprocessively until the enzyme stops 8-11 nucleotides from the 3'-end of the substrate. T4 gene 32 single-stranded DNA-binding protein strongly stimulates the exonuclease activity of T4 RNase H, converting it into a processive nuclease that removes multiple short oligonucleotides with a combined length of 10-50 nucleotides each time it binds to the duplex substrate. 32 protein must bind on single-stranded DNA behind T4 RNase H for processive degradation. T4 RNase H also has a flap endonuclease activity that cuts preferentially on either side of the junction between single- and double-stranded DNA in flap and fork DNA structures. In contrast to the exonuclease, the endonuclease is inhibited completely by 32 protein binding to the single strand of the flap substrate. These results suggest an important role for T4 32 protein in controlling T4 RNase H degradation of RNA primers and adjacent DNA during each lagging strand cycle.

Either bacteriophage T4 RNase H or Escherichia coli DNA polymerase I is essential for phage replication.

Bacteriophage T4 rnh encodes an RNase H that removes ribopentamer primers from nascent DNA chains during synthesis by the T4 multienzyme replication system in vitro (H. C. Hollingsworth and N. G. Nossal, J. Biol. Chem. 266:1888-1897, 1991). This paper demonstrates that either T4 RNase HI or Escherichia coli DNA polymerase I (Pol I) is essential for phage replication. Wild-type T4 phage production was not diminished by the polA12 mutation, which disrupts coordination between the polymerase and the 5'-to-3' nuclease activities of E. coli DNA Pol I, or by an interruption in the gene for E. coli RNase HI. Deleting the C-terminal amino acids 118 to 305 from T4 RNase H reduced phage production to 47% of that of wild-type T4 on a wild-type E. coli host, 10% on an isogenic host defective in RNase H, and less than 0.1% on a polA12 host. The T4 rnh(delta118-305) mutant synthesized DNA at about half the rate of wild-type T4 in the polA12 host. More than 50% of pulse-labelled mutant DNA was in short chains characteristic of Okazaki fragments. Phage production was restored in the nonpermissive host by providing the T4 rnh gene on a plasmid. Thus, T4 RNase H was sufficient to sustain the high rate of T4 DNA synthesis, but E. coli RNase HI and the 5'-to-3' exonuclease of Pol I could substitute to some extent for the T4 enzyme. However, replication was less accurate in the absence of the T4 RNase H, as judged by the increased frequency of acriflavine-resistant mutations after infection of a wild-type host with the T4 rnh (delta118-305) mutant.