The oral microbiota - a mechanistic role for systemic diseases.

Human oral microbiota is the ecological community of commensal, symbiotic, and pathogenic microorganisms found in the oral cavity. Oral microbiota generally exists in the form of a biofilm and plays a crucial role in maintaining oral homeostasis, protecting the oral cavity and preventing disease development. Human oral microbiota has recently become a new focus research for promoting the progress of disease diagnosis, assisting disease treatment, and developing personalised medicines. In this review, the scientific evidence supporting the association that endogenous and exogenous factors (diet, smoking, drinking, socioeconomic status, antibiotics use and pregnancy) modulate oral microbiota. It provides insights into the mechanistic role in which oral microbiota may influence systemic diseases, and summarises the challenges of clinical diagnosis and treatment based on the microbial community information. It provides information for noninvasive diagnosis and helps develop a new paradigm of personalised medicine. All these benefit human health in the post-metagenomics era.

Inhibition of cyclooxygenase-2-mediated matriptase activation contributes to the suppression of prostate cancer cell motility and metastasis.

Chronic inflammation plays an important role in cancer development and progression. Cyclooxygenases-2 (COX-2) is a key enzyme in generating prostaglandins causing inflammation, is often found to be overexpressed in prostate cancer (PCa) and is correlated with PCa cell invasion and metastasis. We aim to investigate the molecular mechanism of how COX-2 promotes PCa cell invasion and metastasis and to evaluate the effect of COX-2 inhibitors in a selected model of PCa progression. Our results showed that the expression of COX-2 and Interleukin 1ß (IL-1ß) was upregulated in highly invasive PCa cells and was correlated with the activated levels of membrane-anchored serine protease matriptase. The expression levels of COX-2 were increased and were correlated with matriptase levels in PCa specimens. Moreover, results showed that COX-2 overexpression or a COX-2 product Prostaglandin E2 (PGE2) caused an increase in matriptase activation and PCa cell invasion, whereas COX-2 silencing antagonized matriptase activation and cell invasion. In addition, the inhibition of COX-2-mediated matriptase activation by Celebrex and sulindac sulfide suppressed the androgen-independent and COX2-overexpressing PCa PC-3 cell invasion, tumor growth and lung metastasis in an orthotopic xenograft model. Our results indicate that COX-2/matriptase signaling contributes to the invasion, tumor growth and metastasis of COX-2-overexpressing and androgen-independent PCa cells.

The study of progesterone action in human myometrial explants.

STUDY HYPOTHESIS: Myometrial explants represent a superior model compared with cell culture models for the study of human myometrial progesterone (P4) signalling in parturition. STUDY FINDING: Gene expression analysis showed myometrial explants closely resemble the in vivo condition and the anti-inflammatory action of P4 is not lost with labour onset. WHAT IS KNOWN ALREADY: Circulating P4 levels decline before the onset of parturition in most animals, but not in humans. This has led to the suggestion that there is a functional withdrawal of P4 action at the myometrial level prior to labour onset. However, to date, no evidence of a loss of P4 function has been provided, with studies hampered by a lack of a physiologically relevant model. STUDY DESIGN, SAMPLES/MATERIALS, METHODS: Myometrial biopsies obtained at Caesarean section were dissected into explants after a portion was immediately snap frozen (t = 0). Microarray analysis was used to compare gene expression of t = 0 with paired (i) explants, (ii) passage 4 myometrial cell cultures or (iii) the hTERT myometrial cell line. Western blotting and chemokine/cytokine assays were used to study P4 signalling in myometrial explants. MAIN RESULTS AND THE ROLE OF CHANCE: Gene expression comparison of t = 0 to the three models demonstrated that explants more closely resemble the in vivo status. At the protein level, explants maintain both P4 receptor (PR) and glucocorticoid receptor (GR) levels versus t = 0 whereas cells only maintain GR levels. Additionally, treatment with 1 µM P4 led to a reduction in interleukin-1 (IL-1) ß-driven cyclooxygenase-2 in explants but not in cells. P4 signalling in explants was PR-mediated and associated with a repression of p65 and c-Jun phosphorylation. Furthermore, the anti-inflammatory action of P4 was maintained after labour onset. LIMITATIONS/REASONS FOR CAUTION: There is evidence of basal inflammation in the myometrial explant model. WIDER IMPLICATIONS OF THE FINDINGS: Myometrial explants constitute a novel model to study P4 signalling in the myometrium and can be used to further elucidate the mechanisms of P4 action in human labour. LARGE SCALE DATA: Data deposited at http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?token=gvmpggkurbgxfqf&acc=GSE77830. STUDY FUNDING AND COMPETING INTEREST: This work was supported by grants from the Joint Research Committee of the Westminster Medical School Research Trust, Borne (No. 1067412-7; a sub-charity of the Chelsea and Westminster Health Charity) and the Imperial NIHR Biomedical Research Centre. The views expressed are those of the author(s) and not necessarily those of the NHS or the Department of Health. The authors have no conflict of interest.

Urethritis in men at the Genito-Urinary Medicine Clinic Kuala Lumpur Hospital.

The clinical features and aetiology of 100 consecutive symptomatic heterosexual male patients with urethritis were studied from March 1994 to August 1994 in the Genito-Urinary Medicine (GUM) Clinic, Kuala Lumpur Hospital. Gonococcal urethritis (GU) was found to be more common (53%) than non-gonococcal urethritis(47%). All patients with GU confirmed microbiologically had clinically evident urethral discharge. Almost half (41%) of the patients with GU developed post-gonococcal urethritis (PGU). The most common organism isolated in PGU was Ureaplasma urealyticum (37%) whilst only 4% had both Chlamydia trachomatis and Ureaplasma urealyticum. Of the 47% of patients with non-gonococcal urethritis (NGU), 50% had no microorganism isolated, 32% had Ureaplasma urealyticum, 7% Chlamydia trachomatis and 11% both Chlamydia trachomatis and Ureaplasma urealyticum.

Negative staining permits 4.0 A resolution with low-dose electron diffraction of catalase crystals.

Low-dose electron diffraction of thin single crystals of catalase that are negatively stained with the light-atom compound, dipotassium glucose-1,6-diphosphate, reveals Bragg reflections extending to 4.0A (= 0.40 nm). Under the same conditions, negative staining with the traditional heavy-metal salt, ammonium molybdate, also gives diffraction spots extending to 4.0 A. These results establish that negative staining of protein crystals preserves periodic structural information into the high-resolution range, unlike the widely accepted current belief that this methodology can give a resolution limited to only 20-25 A.

Molecular phylogeographic studies on Paragonimus westermani in Asia.

The lung fluke, Paragonimus westermani (Kerbert, 1878), is widely distributed in Asia, and exhibits much variation in its biological properties. Previous phylogenetic studies using DNA sequences have demonstrated that samples from north-east Asia form a tight group distinct from samples from south Asia (Philippines, Thailand, Malaysia). Among countries from the latter region, considerable molecular diversity was observed. This was investigated further using additional DNA sequences (partial mitochondrial cytochrome c oxidase subunit 1 (COI) and the second internal transcribed spacer of the nuclear ribosomal gene repeat (ITS2)) from additional samples of P. westermani. Phylogenies inferred from these again found three or four groups within P. westermani, depending on the method of analysis. Populations of P. westermani from north-east Asia use snail hosts of the family Pleuroceridae and differ in other biological properties from populations in south Asia (that use snail hosts of the family Thiaridae). It is considered that the populations we sampled can be divided into two species, one in north-east Asia and the other in south Asia.

Serological evidence of schistosomiasis in the Malaysian police field force.

A total of 1131 Police Field Force personnel were screened serologically for schistosomiasis in Malaysia. A total of 150 (13.3%) were tested positive or borderline. Stool samples from 75 of these cases were however all negative for schistosome eggs. This survey suggests that Police Field Force personnel may be agents for propagating the schistosome life cycle in Malaysia.

The spectrum of cavernous sinus and orbital venous thrombosis: a case and a review.

Orbital venous pathologies encompass a broad range of entities including tumors, shunts, congenital anomalies, aneurysms, and obstructive lesions. Patients may present with a variety of clinical findings which may include a combination of tumefaction, vascular engorgement, orbital pulsation, and exophthalmos, depending on the relationship between the lesion and the vascular system. Clinical findings may be unreliable in excluding serious underlying disorders, and so an extensive clinical and radiologic evaluation is necessary. This article presents a rare case of spontaneous aseptic cavernous sinus-superior ophthalmic vein thrombosis in a woman on hormone replacement therapy, and illustrates the multidisciplinary approach in diagnosis and management. The literature on issues surrounding this case is reviewed.

Effects of mutations in cAMP-dependent protein kinase on chloride efflux in Caco-2 human colonic carcinoma cells.

In order to evaluate the importance of cAMP and cAMP-dependent protein kinase (cAMPdPK) in the regulation of chloride efflux via the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel, Caco-2, human colonic carcinoma cells were transfected with an expression vector encoding a mutant form of regulatory subunit of cAMPdPK under control of the mouse metallothionein 1 promoter. Four stable transformants were isolated that expressed the mutant subunit in a Zn(2+)-inducible manner and exhibited Zn(2+)-inducible inhibition of cAMPdPK activity. The parental and transformed Caco-2 cells were examined for their abilities to regulate chloride efflux in response to various secretagogues using a radioactive iodide-efflux assay. In the transformants, induction of the protein kinase mutation with ZnSO4 markedly decreased chloride efflux in response to forskolin, the 8-(4-chlorophenylthio) analog of cAMP, vasoactive intestinal polypeptide, prostaglandin E2 and isoproterenol, whereas Zn(2+)-treated parental cells remained responsive to these secretagogues. Treatment with carbachol, calcium ionophores or phorbol ester did not acutely affect chloride efflux. Together, these studies indicate that cAMP and cAMPdPK are essential components of secretagogue-regulated chloride channel activity in the Caco-2 cell line. In whole cell patch clamp recordings, induction of the cAMPdPK mutation inhibited anionic conductances indicative of the CFTR chloride channel, whereas purified catalytic subunit of cAMPdPK, added intracellularly, reversed the inhibition. These latter results demonstrate that the CFTR chloride channels in the protein kinase-defective transformants are normal and that the protein kinase mutation specifically affects their regulation, presumably by direct phosphorylation.

Malaysian schistosomiasis: description of a population at risk.

A stable population at risk of Malaysian schistosomiasis was studied. Census results indicated that approximately one-fourth of the inhabitants used a stream where Schistosoma malayensis-infected snails were present as their principal source of water for bathing, drinking, and household tasks. The general population also contacted this stream when fording it or while fishing. Serological surveys using enzyme-linked immunosorbent assay (ELISA) and the circumoval precipitin (COP) test revealed six (9%) and three (4%) positives, respectively, among 67 persons examined. No schistosome ova were found in a general survey of 56 persons which included five ELISA positive and two COP test positive patients. ELISA and COP test prevalences among those dependent on the foci of transmission for water, 13 and 7% respectively, were only slightly higher than prevalences among the remainder of the population, 8 and 4% respectively. These results indicate that even among a stable population at risk of Malaysian schistosomiasis the prevalence is low. Our findings support the hypothesis that S. malayensis is a zoonotic infection in man and that it is unlikely to become a significant public health problem.

Experimental infection of Rattus sabanus and Rattus muelleri with subperiodic Brugia malayi.

R. sabanus and R. muelleri are very common in the lowland forests of Malaysia. In nature they are infected with Breinlia sp. and D. ramachandrani. In an attempt to determine whether they are also susceptible to subperiodic B. malayi and thereby being potential reservoirs of infection of the disease, 24 R. muelleri and 17 R. sabanus were experimentally infected with the parasite. Results show that although they can support the full development of the parasite, they are poor hosts. This confirms the observation that in Malaysia natural infection of Rattus spp. with the parasite has not been seen. These rats therefore are probably not important in the zoonotic transmission of subperiodic B. malayi in Malaysia.