Mode of action and safety of lactosporin, a novel antimicrobial protein produced by Bacillus coagulans ATCC 7050.

AIMS: To determine the mechanism of action of antimicrobial protein, lactosporin, against Gardnerella vaginalis and to evaluate its safety in vitro. METHODS AND RESULTS: Bacillus coagulans ATCC 7050 was grown at 37°C for 18 h. The cell-free supernatant was concentrated 10-fold and screened for antimicrobial activity against indicator strain Micrococcus luteus. The mode of action of lactosporin was determined by measuring the potassium release and monitoring the changes in transmembrane potential (Δψ) and transmembrane pH (ΔpH) of the sensitive cells. Lactosporin caused the efflux of potassium ions from M. luteus cells and dissipation of ΔpH in G. vaginalis, while it had no effect on the Δψ. The safety of lactosporin was evaluated by using EpiVaginal(™) ectocervical (VEC-100) tissue model. Over 80% of the cells in the vaginal tissue remained viable after exposure to lactosporin for 24 h. CONCLUSIONS: Lactosporin potentially exerts its antimicrobial activity by selective dissipation of ΔpH and/or by causing leakage of ions from the sensitive cells. Safety studies suggest that lactosporin is a noncytotoxic antimicrobial for vaginal application. SIGNIFICANCE AND IMPACT OF THE STUDY: This study revealed that lactosporin is an effective and safe antimicrobial preparation with potential application for the control of bacterial vaginosis.

Pulmonary zygomycosis with Cunninghamella bertholletiae in a killer whale (Orcinus orca).

An adult female killer whale (Orcinus orca) was transported to the Port of Nagoya public aquarium in June 2010. While the animal was being maintained in the aquarium there was a gradual decrease in body weight. On October 1st, 2010 the whale exhibited signs of gastrointestinal disease and died on January 14th, 2011. At necropsy examination the gastric compartments were filled with a large number of variably-sized rocks (total weight 81.4 kg) and there was marked ulceration in the third compartment. There were multifocal tubercle-like nodules within the lungs and on sectioning there were numerous abscesses and pulmonary cavities. Microscopically, there was severe suppurative pneumonia associated with fungal hyphae that were infrequently septate and often branched. Numerous bacterial colonies were also present. The hyphae demonstrated immunohistochemical cross-reactivity with Rhizomucor spp. and Cunninghamella bertholletiae was cultured. Bacteriological culture revealed the presence of Proteus mirabilis, Pseudomonas aeruginosa and Pseudomonas oryzihabitans. This case represents the first documentation of zygomycosis associated with C. bertholletiae in a marine mammal.

NATURAL ANTIMICROBIALS AND THEIR ROLE IN VAGINAL HEALTH: A SHORT REVIEW.

Lactobacillus species maintain the vaginal ecosystem in a healthy condition by production of antimicrobial substances. Depletion of lactobacilli in the vagina results in bacterial vaginosis (BV), where the normal flora is replaced by several bacterial pathogens, usually Gardnerella vaginalis and obligate anaerobes. BV may cause complications such as premature labor, low birth weight and increased risk of HIV acquisition. The currently recommended antibiotic treatments for BV are not always effective and often lead to reoccurrence of the infection. In many cases, this is due to the antibiotic-resistant forms of the pathogens. Therefore, there is an interest in the development of treatments using antimicrobials derived primarily from Lactobacillus spp., such as ribosomally produced antimicrobial peptides (bacteriocins) and lactic acid. These substances effectively inhibit pathogenic bacteria, are safe and do not pose any threat to healthy vaginal Lactobacillus spp. It may be possible to find an effective treatment against BV while reducing the infection's reoccurrence and the treatment-related complications through hurdle technology. This would be achieved by combining antimicrobials produced by Lactobacillus spp. with different natural antimicrobials obtained from plants or other non-pathogenic organisms.

Dental otalgia.

BACKGROUND: Secondary or referred otalgia can represent a diagnostic challenge to the otolaryngologist. Collectively, dental disorders are the most common causes of secondary otalgia presenting to the ENT clinic, and may account for up to 50 per cent of referred otalgia. Temporomandibular joint dysfunction syndrome represents the most common dental cause of referred otalgia. Decay and pulpal inflammation of posterior teeth can also frequently present as otalgia. The common embryological developmental origin of both oral and dental structures and the ear is responsible for their overlapping sensory nerve supplies, and this explains referred otalgia secondary to dental and temporomandibular joint disorders. These disorders can be easily overlooked, resulting in unnecessary and costly investigations. AIMS: This review aims to give a succinct overview of common dental causes of otalgia, and to provide ENT clinicians with guidelines for performing a rapid and simple dental and temporomandibular joint examination, which will reliably diagnose or exclude dental otalgia.

Immediate CT pneumocolon for failed colonoscopy; comparison with routine pneumocolon.

AIM: To assess the role and reliability of 2D CT pneumocolon in the diagnosis of colonic malignancy, and compare feasibility of referral sources. MATERIALS AND METHODS: A prospective study of 50 patients with suspected large bowel malignancy. Patients underwent bowel cleansing, rectal air insufflation and contrast enhanced CT with 5 mm collimation, 3 mm reconstruction and a pitch of 1.4. Subsequent correlation was with pathology (16), colonoscopy (13), barium enema (5), ERCP (1) and clinical follow-up alone (8). RESULTS: Diagnostic images were obtained in 43/50 patients (86% feasibility). Follow-up was obtained in 35/43 patients (one patient died of an unrelated cause, and seven patients were deemed unfit for further investigation). Seventeen colonic carcinomas were diagnosed (three false-positives: one ischaemic colitis, one diverticular stricture and one faecal mass), one diverticular stricture, one fistula, one pancreatic carcinoma and one ovarian malignancy. The remaining 14 were negative. Overall sensitivity was 100% (for lesions >1.5 cm) with a specificity of 94% for structural abnormalities, but only 82% for the correct identification of malignancy. CONCLUSION: Computed tomography (CT) pneumocolon is a reliable alternative to barium enema where colonoscopy is incomplete, with the advantage of extraluminal screening, and examination of the proximal bowel. In the frail elderly or young unfit patient, it is a valuable additional diagnostic tool.

Multiple mitochondrial viruses in an isolate of the Dutch Elm disease fungus Ophiostoma novo-ulmi.

The nucleotide sequences of three mitochondrial virus double-stranded (ds) RNAs, RNA-4 (2599 nucleotides), RNA-5 (2474 nucleotides), and RNA-6 (2343 nucleotides), in a diseased isolate Log1/3-8d2 (Ld) of the Dutch elm disease fungus Ophiostoma novo-ulmi have been determined. All these RNAs are A-U-rich (71-73% A + U residues). Using the fungal mitochondrial genetic code in which UGA codes for tryptophan, the positive-strand of each of RNAs 4, 5, and 6 contains a single open reading frame (ORF) with the potential to encode a protein of 783, 729, and 695 amino acids, respectively, all of which contain conserved motifs characteristic of RNA-dependent RNA polymerases (RdRps). Sequence comparisons showed that these RNAs are related to each other and to a previously characterized RNA, RNA-3a, from the same O. novo-ulmi isolate, especially within the RdRp-like motifs. However, the overall RNA nucleotide and RdRp amino acid sequence identities were relatively low (43-55% and 20-32%, respectively). The 5'- and 3'-terminal sequences of these RNAs are different, but they can all be folded into potentially stable stem-loop structures. Those of RNA-4 and RNA-6 have inverted complementarity, potentially forming panhandle structures. Their molecular and biological properties indicate that RNAs 3a, 4, 5, and 6 are the genomes of four different viruses, which replicate independently in the same cell. These four viruses are also related to a mitochondrial RNA virus from another fungus, Cryphonectria parasitica, recently designated the type species of the Mitovirus genus of the Narnaviridae family, and to a virus from the fungus Rhizoctonia solani. It is proposed that the four O. novo-ulmi mitochondrial viruses are assigned to the Mitovirus genus and designated O. novo-ulmi mitovirus (OnuMV) 3a-Ld, 4-Ld, 5-Ld, and 6-Ld, respectively. Northern blot analysis indicated that O. novo-ulmi Ld nucleic acid extracts contain more single-stranded (ss, positive-stranded) RNA than dsRNA for all three newly described mitoviruses. O. novo-ulmi RNA-7, previously believed to be a satellite-like RNA, is shown to be a defective RNA, derived from OnuMV4-Ld RNA by multiple internal deletions. OnuMV4-Ld is therefore the helper virus for the replication of both RNA-7 and another defective RNA, RNA-10. Sequence comparisons indicate that RNA-10 could be derived from RNA-7, as previously suggested, or derived directly from RNA-4.

A potential model for early stages of chromosomal evolution via concentric Robertsonian fans: a large area of polymorphism in southern short-tailed shrews (Blarina carolinensis).

Western Tennessee contains unusually highly polymorphic populations of southern short-tailed shrews (Blarina carolinensis). We previously documented eight Robertsonian translocations (ROBs) accounting for a variation in diploid number from 46 in most of this species' range to 34-40 in western Tennessee. We have now expanded our study to include data from adjacent areas in Tennessee and Mississippi, 10 localities in all. The new data show a variation in diploid number ranging from 31 to 41, four new ROBs (for a total of 12), and the novel finding of monobrachial translocations in this group. All animals collected from this large area (extending over 12, 000 km(2)) had some level of ROBs, and none represented the 2n = 46 form seen in other parts of the range of this species. Because other species of shrews (genus Sorex) are not affected in the same area, the factors and/or selective forces causing this extensive polymorphism in B. carolinensis must be unique to this species and to this geographic area. Some ROBs were found throughout this large area of over 12,000 km(2). Other translocations (including those with monobrachial homology) were located in one or two localities in this large area, and still other translocations were intermediate in their distribution. There was a concentric pattern to the evolution and presumed spreading of the ROBs. This allowed us to expand the concept of a Robertsonian "fan," introduced by Matthey (1970), to that of concentric evolution of multiple fusion fans: ROBs likely arose independently, separated temporally and geographically, and radiated into surrounding populations to create this complex zone of polymorphism. This is an active process in its infancy, and it is not as mature as that seen in European studies of Mus and Sorex.

Emotional and semantic priming as a measure of information processing in young people with school refusal: a research note.

We investigated semantic and emotional priming of lexical decision-making in 20 school refusers and 20 attenders aged 11-16 years, matched for sex and reading ability. We hypothesised that: semantic and emotional priming would be demonstrable in both samples; and that the school refusers would show emotional priming of school-related words to aversive primes. Both samples showed semantic priming; emotional priming was shown by the attenders and 11 school refusers without a history of depression. School-refusing children did not show emotional priming for school-related words. Nine school-refusing children with either current or past depression showed a general reduction in their priming. These results show that both semantic and emotional priming can be detected in this age-range. They do not support school refusal being typically associated with anxiety about school. The reduction in priming in those with a depressive history is similar to inhibition in information processing in depressed adults. Priming is therefore sensitive to at least some psychiatrically relevant states or traits in this age-range. It is concluded that priming could be a useful measure of information processing in this age-group, and further research is warranted.

Fetal monitoring--midwifery attitudes.

OBJECTIVE: to survey midwives' attitudes and practices related to intrapartum fetal monitoring. DESIGN: descriptive correlational study. SETTING: regional and district maternity unit and related community area within one health authority. PARTICIPANTS: all midwives were invited to participate. Two hundred and forty two questionnaires were administered and 117 were returned (48% response rate). MEASUREMENTS AND FINDINGS: in the questionnaire information was collected on professional/demographic details, education and practices related to intrapartum fetal monitoring, together with a 20-item attitude scale which encompassed attitudes towards fetal monitoring and related issues. As expected, the findings suggest that midwives' preferred methods of fetal monitoring varied with the client's risk category. However, midwife preference did not necessarily match actual choice of method. There are many factors influencing choice, not least of which is confidence in ability. Significant differences were found between midwives. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: the findings highlight some of the issues relating to individual confidence. 97% of the midwives felt they would benefit from in-service training in CTG interpretation. The findings support the development of continuing in service education programmes for midwives.

Low prevalence of Helicobacter pylori in inflammatory bowel disease: association with sulphasalazine.

The prevalence of IgG antibodies to Helicobacter pylori was examined in 110 patients with inflammatory bowel disease (IBD) (63 ulcerative colitis, 47 Crohn's disease) and compared with 100 age and sex matched control patients. The overall prevalence of H pylori seropositivity in the IBD patients was 22%, which was significantly less than that of 52% in the controls (p < 0.002). There was no difference in prevalence between ulcerative colitis and Crohn's patients. The low seropositivity in the IBD patients resulted from a very low prevalence of 10% in those currently receiving sulphasalazine (n = 40) and similarly low prevalence of 7% in those previously receiving sulphasalazine (n = 30). In those receiving olsalazine or mesalazine and who had never had sulphasalazine, the prevalence of seropositivity was 45%. Further studies using 14C urea breath test and microscopy of antral biopsy specimens confirmed that the negative serology in patients receiving sulphasalazine resulted from absence of the infection rather than absence of humoral immune response to it. In six control patients with H pylori infection, a two week course of sulphasalazine (500 mg four times daily) only caused slight suppression of the 14C urea breath test. In vitro studies failed to show any direct antibacterial effect of sulphasalazine on H pylori. These findings indicate that longterm treatment with sulphasalazine leads to eradication of H pylori infection and that this does not result from a direct antibacterial effect. It may be caused by the drug treating the gastritis and thereby depriving the bacterium of essential nutrients exuded by the inflamed mucosa.

Safety of general anaesthesia and surgery in acute hepatic porphyria.

Patients with acute hepatic porphyria are denied essential operations because of concern that general anaesthesia and surgery will precipitate a life threatening porphyric crisis. This study assessed the safety of surgery under general anaesthesia in these patients. A combined prospective and retrospective case note study, with a biochemical study, was conducted in 25 patients with acute hepatic porphyria undergoing 38 surgical operations. Clinical outcome measures were survival and occurrence of porphyric crisis after surgery. The biochemical activity of porphyria was assessed by measurement of the perioperative 24 hour excretion of the haem precursors delta amino-laevulinic acid (ALA) and porphobilinogen (PBG). There were no deaths or crises after 29 operations in 19 patients who were known to have porphyria before their surgery, and therefore given only appropriate drugs. These operations include such major procedures as mitral valve replacement, hip replacement, coronary artery grafting, cholecystectomies, and renal transplantation. In eight of these patients the urinary excretion of ALA and PBG were studied, and showed no sustained postoperative increase. Nine operations were performed in eight patients before the diagnosis of porphyria was known and who thus received routine anaesthetic agents. Seven of these patients developed a postoperative porphyric crisis. Two of them died. It is concluded therefore that even the most major surgery can be undertaken safely in patients with porphyria. The risk is for undiagnosed cases.

Parental psychiatric disorder: clinical prevalence and effects on default from treatment.

In a retrospective case note study, 36% of 253 consecutive cases seen and treated at a community child and adolescent clinic had at least one caretaker with a concurrent psychiatric disorder. The most common diagnosis in caretakers was depression (20% of the sample). Both, psychiatric disorder in general, and depression in particular, were significantly related to default from treatment. The relevance of these results to all aspects of child health care is discussed. It is concluded that more research into the impact of parental psychiatric disorder on the treatment of children across various clinical settings is needed.

Haem precursor effects on [3H]-PK 11195 binding to platelets.

The haem precursors delta-aminolaevulinic acid (ALA) and porphobilinogen (PBG), over-produced in acute intermittent porphyria (AIP), may mediate porphyric neuropathy. Porphyrins bind to peripheral benzodiazepine receptors, putative mitochondrial porphyrin translocation sites. In AIP, ALA and PBG may interfere with porphyrin transport causing a deficit in essential haemoproteins. The effects of ALA, PBG and protoporphyrin 1X on the binding of [3H]-PK 11195 to platelets were examined. Ligand binding was also examined in platelets from porphyric patients. The haem precursor protoporphyrin 1X (10 microM) caused an increase in Kd values (p < 0.05) whereas Bmax values remained unaltered. Neither ALA (10 microM) nor PBG (10 microM) altered ligand binding. Patient platelets showed no change in ligand binding values. ALA and PBG are unlikely to compete with porphyrins for peripheral benzodiazepine sites.