Screening of traditional herbal medicine: first, do a retrospective study, with correlation between diverse treatments used and reported patient outcome.
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Humans↗
Biomedical subjects
M Willcox
Publications and source records attributed to M Willcox.
Lipid, lipase and lipocalin differences between tolerant and intolerant contact lens wearers.
PURPOSE: Tear volume is reduced in symptomatic contact lens wearers, evaporation of the ocular tear film may be a cause. In this study we have focussed on symptomatic or intolerant subjects and compared their tear film lipid-related features to those tolerant to soft contact lens wear. METHOD: Fourteen tolerant and 10 intolerant to lens wear subjects were recruited for this study. Intolerance to lens wear was defined as experiencing dryness symptoms in the first 6 hours of lens wear and consequently not being regular lens wearers. Lipid layer appearance was graded on a 0-5 scale, meibomian gland obstruction was observed, and the McMonnies questionnaire completed. Tears were collected without reflex stimulation. Degraded lipid (tear aldehyde content), secretory phospholipase A2 enzyme (sPLA2) concentration and activity and lipocalin concentration were analysed using spectrophotometry to quantify colour reactions and enzyme linked immunosorbent assays. Statistical results were calculated using non-parametric tests (median +/- interquartile range) or chi-squared test. RESULTS: Degradation of polyunsaturated fatty acids and related esters leads to the by-products, malondialdehyde and 4-hydroxy-2(E)-nonenal. Intolerant subjects were found to have significantly (p = 0.004) higher concentrations of these by-products in their tears (0.85 +/- 1.0 microM; n = 9) compared to tolerant subjects (0.15 +/- 0.15 microM; n = 10). Intolerant subjects (1.86 +/- 0.05 ng/microl; n = 9) had significantly more (p = 0.047) sPLA2 enzyme in their tears compared with tolerant subjects (1.80 +/- 0.08 ng/microl; n = 12) and significantly more enzyme activity (p = 0.012). Intolerant subjects had significantly higher amounts of lipocalin in their tears (2.40 +/- 1.5 microg/microl; n = 10, p < 0.001) compared to tolerant subjects (0.45 +/- 0.85 microg/microl; n = 13). CONCLUSION: Changes to the components of the tear film, however small, can disturb the nature and dynamics of the tear film. Increased lipases, degraded lipids and lipocalins in the aqueous tear film potentiates intolerance to contact lens wear and was associated with increased McMonnies dry eye history scores and symptoms scores.
Adult↗
Enzymatic, clinical and histologic evaluation of corneal tissues in experimental fungal keratitis in rabbits.
Mycotic keratitis, being frequently refractive to most of the currently available antifungal therapy, continues to pose a therapeutic challenge to the clinician. In keratitis of infectious etiology stromal dissolution may be brought about by a combination of agent and host factors. An understanding of the source and nature of corneal tissue damage is essential for evolving more effective therapeutic modalities in the treatment of fungal keratitis. In the present study, we have characterized the extracellular proteases produced in vitro by corneal fungal pathogens namely the Aspergillus flavus and Fusarium solani when collagen was provided as the sole nitrogen source. In addition, fungal infected rabbit corneas were investigated for proteolytic activities and nature of inflammatory reaction. Gelatin zymography detected protease bands with molecular mass ranging from 100 to 200 kDa in the culture extracts of A. flavus, and a single major band of molecular mass approximately 200 kDa in the culture extracts of F. solani. A basal proteolytic activity of mass 65 kDa was visualized in all uninfected and infected rabbit corneal extracts. Infected corneas in addition revealed the presence of additional proteolytic species of mass 92 and 200 kDa. The enzyme inhibitory profile suggested that fungal cultures in vitro contained predominantly serine protease activity and to a lesser extent metalloprotease activity. However, fungal infected corneal homogenates showed the presence of metalloproteinase activity alone, the enzymatic activities entirely being sensitive to ethylene diamine tetra acetate (EDTA), a metalloprotease inhibitor. Interestingly, the serine proteolytic activity detected in fungal cultures in vitro was not present in the fungal infected corneas in vivo. However, the possible role of fungal serine proteases in the activation of corneal matrix metalloproteinases (MMPs) cannot be ruled out. Based on the criteria of molecular mass, proteolytic activity in the presence of calcium at neutral pH, and sensitivity to inhibition by a metalloprotease inhibitor, the 65 and 92 kDa gelatinases were identified as MMP 2 and MMP 9, respectively. The expression of 92 and 200 kDa gelatinases correlated positively with the amount of polymorphonuclear cells present in the infected tissues. Activated resident corneal cells or inflammatory cells may largely contribute to the increased proteolytic activities in fungal infected corneas resulting in tissue matrix degradation in fungal keratitis.
Animals↗
The role of IL-1beta in the regulation of IL-8 and IL-6 in human corneal epithelial cells during Pseudomonas aeruginosa colonization.
PURPOSE: Recent studies have shown that the levels of interleukin (IL)-1beta, IL-6 and IL-8 are associated with the severity of infectious diseases. The purpose of this study was to investigate whether IL-1beta regulates the expression of IL-6 and IL-8 in human corneal epithelial cells during Pseudomonas aeruginosa colonization. METHODS: Confluent immortalized human corneal epithelial cells were challenged with P. aeruginosa 6294 in the presence of anti-human IL-1beta antibody or matched control antibody. The cells were also challenged with recombinant IL-1beta protein without bacterial colonization. Expression of IL-1beta, IL-6 and IL-8 mRNA and protein was detected by reverse transcription (RT)-PCR and by enzyme-linked immunosorbent assay (ELISA), respectively. IL-1beta localization was determined by immunohistochemistry. RESULTS: Human corneal epithelial cells expressed low levels of IL-1beta and high levels of IL-6 and IL-8 during P. aeruginosa colonization. Addition of IL-1beta Ab resulted in a significant decrease (P < 0.05) in IL-8 protein expression at 4 h, 8 h and 12 h. Addition of IL-1beta Ab reduced IL-6 protein expression at 8 h and increased IL-6 protein expression at 12 h. Addition of recombinant IL-1beta protein alone strongly stimulated the expression of IL-8 and IL-6. Immunohistochemical staining showed that IL-1beta protein was present both intracellularly and extracellularly in P. aeruginosa colonized cells. CONCLUSIONS: IL-1beta is able to modulate expression of both IL-6 and IL-8 at transcriptional and post-transcriptional levels in human corneal epithelial cells.
Cells, Cultured↗
Effects of exogenous interleukin-6 during Pseudomonas aeruginosa corneal infection.
Lack of interleukin-6 (IL-6) during Pseudomonas aeruginosa corneal infection leads to more severe disease with changes in neutrophil recruitment. Exogenous IL-6 leads to increased efficiency of neutrophil recruitment and reduced bacterial loads in corneal infection in both IL-6 gene knockout and wild-type mice. This may be mediated by IL-6 increasing the production of corneal macrophage inflammatory protein 2 and intercellular cell adhesion molecule 1. We conclude that effective recruitment of neutrophils into the cornea is dependent on the production of IL-6 and that early augmentation of IL-6 may be protective in corneal infection.
Animals↗
New research initiative on plant-based antimalarials.
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Antimalarials↗
Towards a closed eye model of the pre-ocular tear layer.
Although the tear film has been extensively studied as it exists in the open eye state, until recently very little was known as to what happens to the tear film on eye closure. Recent studies have shown that eye closure results in a profound change in the composition, origins, turnover and physiological functions of the tear film. These changes include a shift from an inducible, neurologically controlled, lacrimal secretion containing among other proteins primarily lysozyme, lactoferrin and tear specific lipocalin, to a much slower, constitutive-type of secretion, composed almost exclusively of sIgA. This change is accompanied by the build-up of sialoglycoproteins of epithelial and goblet cell origin, the build-up and activation of complement and the build-up of serum proteins. In addition, various cytokines and proinflammatory mediators accumulate, including some which are potent inducers of angiogenesis and leukochemotaxis. The closed eye also exhibits the recruitment and activation of massive numbers of PMN cells. This results in a stagnant, closed eye layer, which is extremely rich in reactive complement products, PMN cell proteases including protease-3, elastase, capthepsin G, MMP-9 and urokinase. We have postulated that this shift represents a fundamental change in host-defense strategies from a passive-barrier defense to an active immune, inflammatory, phagocyte-mediated process and that this shift is necessitated in order to protect the cornea from entrapped microorganisms. Studies have shown that autologous cell damage is avoided in closed eye tear fluid, by the accumulation of several modulators of complement activation, which shift activation towards opsonization of entrapped microorganisms and the build-up of a wide array of antiproteases. Some of the latter are likely to arise from the ocular surface tissues. Corneal neovascularization may be avoided in part by the build-up of alpha2-macroglobulin and the conversion of plasminogen to angiostatin. It is highly probable that other bioactive protein fragments are produced in the closed eye, which contribute to homeostasis. Areas of future study are indicated.
Circadian Rhythm↗
KC production in the cornea in response to Pseudomonas aeruginosa challenge.
Pseudomonas aeruginosa can cause ulcerative bacterial keratitis. A feature of keratitis is the rapid infiltration of the avascular corneal stroma by neutrophils. KC is a potent neutrophil chemokine. The present study used a mouse model of ocular infection to assess the relationship between KC and inflammation in the cornea in response to challenge with a strain of P. aeruginosa causing keratitis. Low levels of KC mRNA and protein were detected by in situ hybridization and ELISA, respectively, in unchallenged corneas. Dramatically increased numbers of KC mRNA+ cells were present in P. aeruginosa strain 6294-challenged corneas. Expression of KC mRNA was found to be up-regulated in the corneal epithelium in response to wounding alone. The KC mRNA+ cells were located in the epithelium and corresponding to infiltrating neutrophils cells in the stroma. Quantification of KC protein at different time points showed peak levels at 8 h of bacterial challenge. These results suggest that KC may be involved with the regulation of leucocyte infiltration early during bacterial keratitis.
Animals↗
Differential interleukin-6 mRNA expression in Nippostrongylus brasiliensis infection of susceptible and resistant strains of mice.
Intestinal parasitic infection is still a major problem in humans and animals, yet host immunity against gut parasitic infection remains partially understood. Eosinophilia and mastocytosis are features of such infection that have been shown to be genetically controlled. The expression of IL-6 is detected in eosinophils, mast cells and neutrophils and may be responsible for the regulation of leucocytes at infective sites. The relationships between IL-6 expression, eosinophilia, mastocytosis and host immunity remain unclear. In the present report, a close correlation between IL-6 mRNA+ cells, eosinophilia, mastocytosis and worm expulsion is demonstrated, which may indicate a role for IL-6 in regulation of host immunity against intestinal parasite infection.
Animals↗
The differential regulation of nitric oxide by Herpes simplex virus-1 and -2 in a corneal epithelial cell line.
Herpes simplex keratitis s a significant cause of blindness worldwide. Nitric oxide (NO) has been shown to play a role in non-specific defence mechanisms and cell signalling in bacterial and parasitic infections. We investigated if Herpes simplex virus (HSV) isolated from keratitis could induce NO production. Human corneal epithelial cells were infected with high (multiplicity of infection; MOI 0.4) and low (MOI 0.04) HSV-1 and HSV-2 concentrations. Culture supernatants were collected at 1 h, 4 h, 8 h, 12 h and 24 h postchallenge. Samples were prepared by removal of proteins by ultrafiltration. Production of NO was measured using nitrite and nitrate assays. Herpes simplex virus-1 downregulated the production of NO, while HSV-2 upregulated NO production. Downregulation of NO could be a survival strategy against the cytotoxic action of NO, to eliminate infected cells. Upregulation of NO production may be associated with the presence of glycoproteins on the viral coat, which have been shown to induce NO in other disease conditions. Further studies are required to confirm the role of NO in viral keratitis.
Adenoviridae↗
The first international meeting of the Research Initiative on Traditional Antimalarial Methods (RITAM).
The first international meeting of the Research Initiative on Traditional Antimalarial Methods (RITAM) was held at the Regional Dermatology Training Centre (RDTC) of the Tumaini University of Health Sciences, Moshi, Tanzania, on December 8-11, 1999. This Inaugural Meeting of RITAM, jointly hosted by the Global Initiative for Traditional Systems of Health (GIFTS) at Oxford University and the World Health Organization (WHO), was designed to develop a strategy for more effective, evidence-based use of traditional medicines that can also inform malaria-control policy decisions. RITAM was established during 1999 as a network of researchers and other people who are active or interested in the study and use of traditional, plant-based antimalarials. RITAM is a partnership between GIFTS of Health, University of Oxford and the Tropical Disease Research (TDR) Programme of WHO. Malaria is one of the key health issues affecting developing countries, particularly in sub-Saharan Africa and Asia. With increasing drug resistance and the high cost of pharmaceutical drugs, the use of herbal antimalarials is popular. The conference was attended by biologic and social scientists, clinicians, traditional healers, and policy makers from Africa, Asia, Europe, and the Americas. The meeting was funded by the Rockefeller Foundation, the Nuffield Foundation's Commonwealth Programme, WHO's TDR Programme, and direct support to delegates was provided by other funders. The meeting addressed the need for research and policy on the prophylactic and therapeutic effects of medicinal plants as well as on vector control and repellence. There were five main outputs from the meeting: (1) targets for making a significant contribution to the control of malaria through the use of traditional antimalarial methods; (2) methods for achieving these targets, including ethical guidelines; (3) an implementation strategy for moving this field ahead quickly and soundly and for putting research findings into practice; (4) linkages established between researchers working on traditional antimalarial methods, based on agreed research priorities and designed to avoid unnecessary replication; and (5) strengthening the RITAM database of current knowledge on traditional herbal antimalarial methods. Four specialist groups were established to develop the above: (1) policy, advocacy, and funding; (2) preclinical studies; (3) clinical development; and (4) repellance and vector control. These will be coordinated by an executive committee managed by GIFTS. Two meetings are planned in 2000: a natural-products chemistry meeting at WHO in Geneva, Switzerland, in June; and a symposium at the World Congress on Tropical Medicine in Cartagena, Colombia, in August.
Antimalarials↗
Infiltrative keratitis associated with extended wear of hydrogel lenses and Abiotrophia defectiva.
PURPOSE: Infiltrative keratitis is a common complication associated with extended wear of hydrogel lenses. Causative bacteria are often isolated from the lens at the time of an event. We report a case where three repeated occurrences of infiltrative keratitis were associated with contamination of the contact lenses by Abiotrophia defectiva. METHODS: A 34-year-old man participating in a clinical trial of extended wear hydrogel contact lenses experienced three episodes of infiltrative keratitis. The clinical presentation was observed using a biomicroscope. At the time of each event, the contact lenses were removed aseptically and ocular swabs were taken for bacterial identification and enumeration. The condition was monitored until full resolution. RESULTS: The condition was characterized by irritation, marked bulbar and limbal injection, and multiple focal subepithelial infiltrates. Many of the infiltrates also showed overlying staining with fluorescein. In each of the three events of infiltrative keratitis, A. defectiva was cultured from the contact lens and ocular swabs. CONCLUSION: This is the first reported occurrence of infiltrative keratitis associated with A. defectiva contamination of contact lenses.
Adult↗
Bacterial colonization of disposable soft contact lenses is greater during corneal infiltrative events than during asymptomatic extended lens wear.
Microorganisms, especially gram-negative bacteria, are considered to play a role in the etiology of certain corneal infiltrative events (CIEs) observed during soft contact lens wear. This study explored the possibility of microbial colonization of soft contact lenses as a risk factor leading to CIEs. In a clinical trial conducted from March 1993 to January 1996, 330 subjects wore disposable soft contact lenses on a 6-night extended-wear and disposal schedule. During this period, 4,321 lenses (118 during CIEs; 4,203 during asymptomatic lens wear) were recovered aseptically and analyzed for microbial colonization. A greater percentage of lenses were free from microbial colonization during asymptomatic wear than during CIEs (42 versus 23%; P < 0.0001). The incidence of gram-positive bacteria, gram-negative bacteria and fungi was greater during CIEs than during asymptomatic lens wear (P < 0.05). During asymptomatic lens wear, gram-positive bacteria were isolated most frequently and were usually normal external ocular microbiota. Of the gram-positive bacteria, the incidence of Streptococcus pneumoniae was greater during CIE than during asymptomatic wear (7.6 versus 0.6%; P < 0. 0001). While gram-negative bacteria were seen in few cases during asymptomatic wear, their incidence during CIE in comparison to asymptomatic wear was substantial and significant (23.7 versus 3.8%; P < 0.0001). Also, the level of colonization was high. Of CIEs, events of microbial keratitis, contact lens acute red eye, and asymptomatic infiltrative keratitis were associated with lens colonization with gram-negative bacteria or S. pneumoniae. Colonization of soft contact lenses with pathogenic bacteria, especially gram-negative bacteria and S. pneumoniae, appears to be a significant risk factor leading to CIE.
Bacteria↗
Fair trade for less developed countries?
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Commerce↗
TNF-alpha production in the cornea in response to Pseudomonas aeruginosa challenge.
Pseudomonas aeruginosa can cause ulcerative bacterial keratitis or contact lens-induced acute red eye (CLARE) in humans. The present study used a mouse model of ocular infection and inflammation to examine the relationship between TNF-alpha and inflammation in the cornea in response to challenge with either a strain of P. aeruginosa causing keratitis or a CLARE strain. Constitutive TNF-alpha mRNA was detected in the epithelium, mainly towards the periphery. After infection with the keratitis-inducing strain (6294), TNF-alpha expression was elevated four-fold by 24 h post-challenge. No detectable induction of TNF-alpha mRNA was seen with CLARE strain (Paer1) challenge at any time point. The TNF-alpha protein production detected by ELISA showed a corresponding pattern to the mRNA expression, which also correlated with pathological changes. These results suggest that invasive strains of P. aeruginosa create greater pathological changes as a result of elevated TNF-alpha production, which contributes to inflammation during keratitis in vivo.
Animals↗
Colonization of hydrogel lenses with Streptococcus pneumoniae: risk of development of corneal infiltrates.
PURPOSE: To report the association between colonization of contact lenses with Streptococcus pneumoniae and the observation of corneal infiltrative events in a group of patients wearing disposable hydrogel lenses on an extended-wear schedule. METHODS: In a prospective clinical trial, 330 patients wore disposable hydrogels on a 6-night extended-wear and replacement schedule. The contact lens, lid, and conjunctival microbiota of these subjects was analyzed at frequent intervals and at the time of an adverse event. RESULTS: Streptococcus pneumoniae was an uncommon isolate and was recovered from only one of the 3,763 conjunctival samples, five (0.1%) of the 3,764 lid samples, and 33 (0.8%) of the 4,315 contact lens samples. Of the 33 lens samples, 10 (30%) were associated with corneal infiltrative events. Many of the events were mild inflammatory responses and resolved rapidly on discontinuation of lens wear. The presence of S. pneumoniae on the contact lens was associated with a significant risk of development of corneal infiltrates (odds ratio, 3.0; p = 0.0227, logistic-regression analysis). CONCLUSIONS: Presence of S. pneumoniae on hydrogel lenses is a significant risk factor for the development of corneal infiltrates.
Adolescent↗
Expression of interleukin-6 in the cornea in response to infection with different strains of Pseudomonas aeruginosa.
Strains of Pseudomonas aeruginosa causing keratitis can be either cytotoxic (6206) or invasive (6294), while a strain (Paer1) causing contact lens-induced acute red eye has been shown to be neither. In situ hybridization was used to examine the location and identity of cells expressing interleukin-6 (IL-6) mRNA in the murine cornea and changes in expression in response to infection with different strains of P. aeruginosa. The number of IL-6-positive cells was determined by image analysis. IL-6 protein levels were measured by an enzyme-linked immunosorbent assay. BALB/c mice were challenged by use of the wounded-cornea model with P. aeruginosa 6294, 6206, or Paer1 (2 x 10(6) CFU). At time intervals up to 24 h, postchallenge corneal tissue was probed for IL-6 mRNA. IL-6 mRNA expression was rapidly elevated in the epithelium in response to strains 6294 and 6206. At the conclusion of the experiments, infiltrating inflammatory cells also stained positively for IL-6 mRNA. In contrast, corneas challenged with strain Paer1 showed significant upregulation of IL-6 mRNA only at 4 h postchallenge. Three distinct patterns of IL-6 mRNA expression in the mouse cornea occur in response to these three ocular isolates of P. aeruginosa. The data obtained for mRNA expression in the cornea for all three strains of P. aeruginosa correlated well with IL-6 protein analysis of whole-eye homogenates. Differences in the cytokine responses to these strains correlate with differences in the pathology associated with each strain and may offer an opportunity to develop strategies for the improved management of ocular inflammation.
Animals↗
Spores of Henneguya salminicola in human stool specimens.
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Animals↗