Evaluation of PCR primers to identify Porphyromonas endodontalis in apical periodontitis clinical samples.

We aimed to evaluate whether two commonly used PCR primers are effective to identify P. endodontalis and discriminate from other prevalent black-pigmented bacteria in apical periodontitis (AP). Endodontic canal samples from patients with asymptomatic AP (n = 20) were collected and cultured in anaerobiosis. Two primer sets to detect P. endodontalis were selected from the literature and first analyzed for their specificity in silico; and then tested on clinical isolates in vitro and finally, in apical exudates ex vivo. The identity of P. endodontalis was verified by PCR and Sanger sequencing with universal primers for bacterial V3-V6 regions 16S rDNA. Only one primer set showed specificity only for P. endodontalis clones in silico and also was specific for P. endodontalis in vitro and ex vivo.

The Salivary Mycobiome Contains 2 Ecologically Distinct Mycotypes.

A broad range of fungi has been detected in molecular surveys of the oral mycobiome. However, knowledge is still lacking on interindividual variability of these communities and the ecologic and clinical significance of oral fungal commensals. In this cross-sectional study, we use internal transcribed spacer 1 amplicon sequencing to evaluate the salivary mycobiome in 59 subjects, 36 of whom were scheduled to receive cancer chemotherapy. Analysis of the broad population structure of fungal communities in the whole cohort identified 2 well-demarcated genus-level community types (mycotypes), with Candida and Malassezia as the main taxa driving cluster partitioning. The Candida mycotype had lower diversity than the Malassezia mycotype and was positively correlated with cancer and steroid use in these subjects, smoking, caries, utilizing a removable prosthesis, and plaque index. Mycotypes were also associated with metabolically distinct bacteria indicative of divergent oral environments, with aciduric species enriched in the Candida mycotype and inflammophilic bacteria increased in the Malassezia mycotype. Similar to their fungal counterparts, coexisting bacterial communities associated with the Candida mycotype showed lower diversity than those associated with the Malassezia mycotype, suggesting that common environmental pressures affected bacteria and fungi. Mycotypes were also seen in an independent cohort of 24 subjects, in which cultivation revealed Malassezia as viable oral mycobiome members, although the low-abundance Malassezia sympodialis was the only Malassezia species recovered. There was a high degree of concordance between the molecular detection and cultivability of Candida, while cultivation showed low sensitivity for detection of the Malassezia mycotype. Overall, our work provides insights into the oral mycobiome landscape, revealing 2 community classes with apparently distinct ecologic constraints and specific associations with coexisting bacteria and clinical parameters. The utility of mycotypes as biomarkers for oral diseases warrants further study.

Maternal periconceptional and first trimester protein restriction in beef heifers: effects on placental parameters and fetal and neonatal calf development.

Few studies have investigated the effects of nutrition during the periconception and early gestation periods on fetal and placental development in cattle. In this study, nulliparous yearling heifers (n=360) were individually fed a diet high or low in protein (HPeri and LPeri) beginning 60 days before conception. From 24 to 98 days after conception, half of each treatment group was changed to the alternative high- or low-protein diet (HPost and LPost) yielding four groups in a 2×2 factorial design. A subset of heifers (n=46) was necropsied at 98 days after conception and fetoplacental development assessed. Placentome number and volume decreased in response to LPeri and LPost diets respectively. Absolute lung, pancreas, septum and ventricle weights decreased in LPost versus HPost fetuses, whereas the post-conception diet altered absolute and relative liver and brain weights depending on sex. Similarly, changes in fetal hepatic gene expression of factors regulating growth, glucose output and lipid metabolism were induced by protein restriction in a sex-specific manner. At term, neonatal calf and placental measures were not different. Protein restriction of heifers during the periconception and early gestation periods alters fetoplacental development and hepatic gene expression. These changes may contribute to functional consequences for progeny, but this may not be apparent from gross morphometry at birth.

Choosing foods for infants: a qualitative study of the factors that influence mothers.

BACKGROUND: Examining the experiences of parents making food choices for infants is important because ultimately this influences what infants eat. Infancy is a critical period when food preferences and eating behaviour begin to develop, shaping dietary patterns, growth and health outcomes. There is limited evidence regarding what or why foods are chosen for infants. OBJECTIVE: To describe the experiences of mothers making food choices for their infant children. METHODS: Semi-structured interviews with 32 Australian mothers of infants aged four to 15 months from a range of socioeconomic backgrounds. An inductive thematic analysis through a process of constant comparison was conducted on transcribed interviews. RESULTS: Mothers described many ideas and circumstances which influenced food choices they made for infants. Themes were developed which encapsulate how the wider environment and individual circumstances combine to result in the food choices made for infants. Beliefs, values, norms and knowledge were a central influence on choices. Cost, quality and availabilities of various foods were also key factors. Related to this, and combined with inherent factors such as perishability and infant acceptability, fresh fruits and vegetables were often singled out as an easy or difficult choice. Influences of time, parents' capacities, social connections and different information sources were clearly apparent. Finally infants' own preferences and how parents helped infants with learning to eat were also key influences on food choices. CONCLUSIONS: Choosing foods for infants is a complex social practice. An ecological framework depicting the multiple influences on what people eat and sociological theory on food choice regarding the role of 'social structure' and 'human agency' are both applicable to the process of choosing foods for infants. Equity issues may be key regarding the degree to which mothers can choose particular foods for infants (e.g. choosing foods which promote health).

The Porphyromonas gingivalis O antigen is required for inhibition of apoptosis in gingival epithelial cells following bacterial infection.

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis infection induces apoptosis inhibition in gingival epithelial cells; however, it is not fully understood which bacterial effectors are involved in this process. The aim of this study is to evaluate whether the P. gingivalis lipopolysaccharide (LPS), specifically the O-antigen region, affects adherence, invasion, viability and apoptosis of gingival epithelial cells. MATERIAL AND METHODS: Gingival epithelial cells (OKF6/TERT2 line) were infected by different freshly prepared P. gingivalis clinical isolates, obtained from subjects with chronic periodontitis (CP3 and CP4) and healthy individuals (H1 and H3). Periodontitis and healthy isolates show differences in O-antigen production, as healthy isolates lack the O-antigen region. In addition, cells were infected by a site-specific mutant lacking the O-antigen portion. After 24 h postinfection, cell proliferation, viability and apoptosis were evaluated by Trypan blue, MTS and annexin V assays, respectively. Bacterial invasion, adhesion and proliferation were measured by gentamicin/metronidazole protection assays. Finally, toll-like receptor (TLR)2 and TLR4 mRNA expression was evaluated by quantitative reverse transcription-polymerase chain reaction. Statistical analysis was performed using ANOVA, Tukey's or Dunnett's tests (p < 0.05). RESULTS: At 24 h postinfection, strains lacking the O-antigen region (healthy isolates and O-antigen ligase-deficient strain) were unable to increase proliferation and viability, or decrease apoptosis as compared with strains producing intact LPS (periodontitis isolates and reference strain). However, the presence of the O-antigen neither contributed to changes in the ability of the bacteria to adhere to or invade cells, nor to intracellular survival. The presence of O-antigen also increased the expression of TLR4 (nearly sixfold), which correlated with inhibition of apoptosis. CONCLUSION: The O-antigen region of P. gingivalis LPS is required to increase gingival epithelial cell viability upon infection by bacteria and this increase is attributable to a reduction in apoptosis. Moreover, although bacterial internalization is required, the effects observed are not due to alterations in P. gingivalis adherence, invasion or intracellular survival. Interestingly, inhibition of apoptosis correlates with increased TLR4 expression, suggesting a role for TLR4 in this process.

An exploration of the views of Australian mothers on promoting child oral health.

BACKGROUND: An important role for parents and caregivers in the prevention of dental caries in children is the early establishment of health promoting behaviours. This study aimed to examine mothers' views on barriers and facilitators to promoting child and family oral health. METHODS: Semi-structured interviews were undertaken with a purposive sample of mothers (n = 32) of young children. Inductive thematic analysis was conducted. RESULTS: Parental knowledge and beliefs, past experiences and child behaviour emerged as major influences on children's oral health. Child temperament and parental time pressures were identified as barriers to good oral health with various strategies reported for dealing with uncooperative children at toothbrushing time. Parental oral health knowledge and beliefs emerged as positive influences on child oral health; however, while most mothers were aware of the common causes of dental caries, very few knew of other risk factors such as bedtime feeding. Parents' own oral health experiences were also seen to positively influence child oral health, regardless of whether these were positive or negative experiences. CONCLUSIONS: Understanding parental oral health beliefs is essential to overcoming barriers and promoting enablers for good child oral health. Improving child oral health also requires consideration of child behaviour, family influences, and increasing awareness of lesser-known influencing factors.

Foraminiferal assemblages in Biscayne Bay, Florida, USA: responses to urban and agricultural influence in a subtropical estuary.

This study assessed foraminiferal assemblages in Biscayne Bay, Florida, a heavily utilized estuary, interpreting changes over the past 65 years and providing a baseline for future comparisons. Analyses of foraminiferal data at the genus level revealed three distinct biotopes. The assemblage from the northern bay was characterized by stress-tolerant taxa, especially Ammonia, present in low abundances ( approximately 2.0 x 10(3) foraminifers/gram) though relatively high diversity ( approximately 19 genera/sample). The southwestern margin of the bay was dominated by Ammonia and Quinqueloculina, an assemblage characterized by the lowest diversities ( approximately 12 genera/sample) and highest abundances ( approximately 1.1 x 10(4) foraminifers/gram), influenced by both reduced salinity and elevated organic-carbon concentrations. A diverse assemblage of smaller miliolids and rotaliids ( approximately 26 genera/sample) characterized the open-bay assemblage, which also had a significant component ( approximately 10%) of taxa that host algal endosymbionts. In the past 65 years, populations of symbiont-bearing taxa, which are indicators of normal-marine conditions, have decreased while stress-tolerant taxa, especially Ammonia spp., have increased in predominance.

A confidential inquiry into asthma deaths in Wales.

BACKGROUND: Death from asthma is regarded as preventable in principle, especially under the age of 65 years. METHODS: In 1994 a confidential inquiry was set up to investigate deaths attributable to asthma in residents of Wales under the age of 65 years. During the period of the inquiry 92 cases were notified as being ascribed to asthma, or (in 1996) having a mention of asthma on the death certificate. Of these, 80 were investigated further with the help of general practitioners, hospital notes, and relatives. The details were then considered by a small panel of doctors who endeavoured to identify factors that may have contributed to the deaths. RESULTS: Asthma was considered to be the underlying cause of 52 deaths. Although disease severity was usually a major factor, some aspect of the patient's behaviour or circumstances seemed to have contributed to 31 deaths, while in 15 cases there was probably a deficiency in medical care. CONCLUSIONS: Some preventable asthma deaths still occur, particularly in relation to inadequate treatment. Factors associated with patients' behaviour and circumstances are more difficult to tackle but, if doctors are aware of high risk patients, increased vigilance may prevent some deaths.

Incidence of psychotic illness in London: comparison of ethnic groups.

OBJECTIVE: To compare annual incidences of psychosis in people from different ethnic groups as defined in the 1991 census. SETTING: Catchment area of district psychiatric hospital. DESIGN: All people aged 16 to 54 years who made contact with a wide range of community and hospital services between 1 July 1991 and 30 June 1992 were screened for psychotic symptoms. Patients with such symptoms were interviewed face to face to collect information on demography, ethnic group, psychiatric history and symptoms, drug use, and how care had been sought. A key informant, usually a close relative, was also interviewed. MAIN OUTCOME MEASURES: Age standardised incidence of schizophrenia and non-affective psychosis according to the ninth edition of the International Classification of Diseases in each ethnic group. RESULTS: Ninety three patients took part, of whom 38 were assigned a certain or very likely diagnosis of schizophrenia (15 in white population, 14 in black, seven in Asian, and two in others). The age standardised annual incidence of schizophrenia was 2.2 (95% confidence interval 1.5 to 2.9) per 10,000 of the population. The incidence ratio for schizophrenia in all ethnic minority groups compared with the white population was 3.6 (1.9 to 7.1); the corresponding figure for non-affective psychosis was 3.7 (2.2 to 6.2). CONCLUSIONS: Raised incidences of schizophrenia were not specific to the African Caribbeans, which suggests that the current focus on schizophrenia in this population is misleading. Members of all ethnic minority groups were more likely to develop a psychosis but not necessarily schizophrenia. The personal and social pressures of belonging to any ethnic minority group in Britain are important determinants in the excess of psychotic disorders found.

Folinic acid and enhanced renal elimination in formic acid intoxication.

Ingestion of over 60 g of formic acid by an adult is potentially fatal. We report a case of a 36-year-old woman with a history of depression who ingested 110 g of formic acid. She survived a complicated intensive care hospitalization following usage of intravenous folinic acid, urinary alkalinization, intravenous furosemide and supportive care. We suggest a management protocol aimed at minimizing formate toxicity by enhancing hepatic formate degradation via the folinic acid 'one carbon pool' and by enhanced renal elimination of formate.

Improved maintenance of remission in ulcerative colitis by balsalazide 4 g/day compared with 2 g/day.

The efficacy of two doses of balsalazide for the maintenance of remission in patients with ulcerative colitis was compared in a double-blind multicentre trial. Sixty-five patients received a 2 g daily dose, and 68 a 4 g dose. The patient groups were similar at entry for sex, age, and disease distribution. Clinical assessment was carried out at 3-monthly intervals, with sigmoidoscopy, rectal biopsy, and blood tests on entry and at 26 and 52 weeks. Clinical relapse over twelve months was significantly less common on the 4 g dose (36%), than on the 2 g dose (55%), P less than 0.01. There were eight withdrawals on 2 g daily and 13 on 4 g daily, six and nine respectively being mainly due to gastrointestinal intolerance. It is concluded that balsalazide is a well-tolerated drug, and is effective for the maintenance of remission in patients with ulcerative colitis, the optimal dose being greater than 2 g daily.

Sedation for endoscopy: midazolam or diazepam and pethidine?

One hundred patients received either diazepam given with pethidine, antagonized with naloxone, or midazolam alone in a double-blind randomized study of sedation for upper gastrointestinal endoscopy. Midazolam produced better amnesia for the procedure (P less than 0.0001) but diazepam and pethidine resulted in less retching during the procedure (P less than 0.01) and less sedation after the procedure, as judged by a simple performance test (P less than 0.02) and patient recall of results (P less than 0.02).

Does ulcerative colitis behave differently in the elderly?

The clinical course of all new patients with ulcerative colitis or indeterminate colitis aged 65 years or older diagnosed in one health district (population 274,000) between 1 January 1975 and 31 December 1984 has been compared with that of all new patients 55 years or younger diagnosed in the same period. Elderly patients were admitted more often in a first attack and were more likely to receive systemic steroids. No elderly patient required urgent surgery and no patient in either group died of ulcerative colitis or its complications. The mortality amongst elderly patients with ulcerative colitis was no greater than expected in a population of the same age. The prognosis for ulcerative colitis in elderly patients is the same as for younger patients.

Surveillance in ulcerative colitis: burdens and benefit.

A review of all patients with ulcerative colitis in one health district between 1975-84 revealed an incidence and prevalence of 7.1 and 84/100,000 population respectively. One hundred and ninety five new patients were diagnosed and 313 patients seen and followed up in the clinic for 1168 patient years. None of these patients died from colitis or a complication. On routine colonoscopy three cases had high grade dysplasia and two asymptomatic carcinomas (Duke's stage A and B). Eighty four patients were known to have ulcerative colitis, but were lost to follow up from the hospital clinic; the total time they were not under hospital surveillance was 315 patient years. At the end of the study these patients were contacted or clinical details obtained from their general practitioners. Five of these patients subsequently presented with symptomatic carcinomas (two Duke's B, one Duke's C and two with metastases); three of these five patients have died from their tumours. Of 48 patients thought to have only mild colitis on initial investigation 21 (43%) had substantial colitis (and two carcinomas) on colonoscopy after eight years of disease. Therefore, patients with apparently distal colitis should be followed in the clinic as well as those with known extensive colitis. For a surveillance programme in a district general hospital, eight patients per 100,000 population need to be seen weekly, 12 colonoscopies/100,000 population need to be carried out annually and the cost for each carcinoma detected is approximately 6015 pounds.

Acute colitis in a district general hospital.

A review of all patients who had been admitted to hospital with acute ulcerative colitis in one health district between 1975 and 1984 showed that 96 had required 114 admissions with acute colitis: 42% (40) were admitted during their first attack, and 20% (19) required urgent surgery. A further nine patients underwent surgery after responding initially to intensive medical treatment that did not check the attack. There were no deaths from acute colitis. Thirteen patients underwent elective surgery for ulcerative colitis, and there were no deaths. The prognosis for acute colitis in district general hospitals has improved.

Sedation for endoscopy: a comparison between diazepam, and diazepam plus pethidine with naloxone reversal.

The combination of diazepam plus pethidine, reversed with naloxone, was compared in a double-blind randomized study with diazepam alone in 100 patients undergoing endoscopy. Patients accepted both methods of sedation equally well. There was no significant difference in sedation after endoscopy, but the addition of pethidine produced a significant improvement in patient co-operation during the endoscopy (P less than 0.001).

A prospective randomized study of effect of proximal gastric vagotomy and vagotomy and antrectomy on bile reflux, endoscopic mucosal abnormalities and gastritis.

71 patients participated in a double-blind trial which compared proximal gastric vagotomy (PVG) with vagotomy and antrectomy (V & A). 82 percent of the patients subsequently volunteered for endoscopy 6 to 12 months after operation and 65 percent for measurement of fasting bile reflux (FBR) and peak acid output (PAO). The results of these follow-up assessments are given in this paper. None of the 36 patients who had undergone V & A had a recurrent ulcer; in contrast ulcers or fresh scars were found in 5 of 35 patients after PGV, even in 2 who had no symptoms. Erythema of the gastric mucosa was seen more commonly after V & A than PGV. Such erythema was associated with high levels of fasting bile reflux (an objective measure of reflux of bile into the stomach) and with symptoms of bile vomiting and mild epigastric pain. High levels of fasting bile reflux were not found after PGV. Histological gastritis of the proximal stomach was equally common after both operations in patients without a recurrent ulcer. Gastritis was not related to endoscopic mucosal erythema or fasting bile reflux, but did correlate with peak acid output. These results confirm that bile reflux is associated with mucosal erythema and symptoms after V & A but that significant bile reflux does not occur after PGV. However, bile reflux is not related to gastritis, which appears to be the result of an operation (either PGV or V & A) which successfully reduces peak acid output.