Inflammatory potential of monospecies biofilm matrix components.

AIM: To assess the inflammatory potential of biofilm matrix constituents of Enterococcus faecalis and Pseudomonas aeruginosa monospecies biofilms on macrophages. METHODOLOGY: In vitro biofilms of E. faecalis and P. aeruginosa were grown (7 days) in aerobic and anaerobic conditions. The biofilm matrix components: exopolysaccharides (EPS) and extracellular DNA (eDNA) were extracted and quantified. The inflammatory potential of EPS and eDNA was assessed on macrophage cell lines (RAW 267.4) using nitric oxide (NO), and enzyme-linked immunosorbent assay for tumour necrosis factor (TNF-α) and interleukin-6 (IL-6) expressions. LPS from P. aeruginosa and planktonic bacteria were positive controls. One-way analysis of variance and the Tukey post hoc test were used for statistical analysis. RESULTS: Extracted EPS from both biofilm strains was associated with higher levels than eDNA in both growth conditions (P < 0.05). The biofilm components had less inflammatory potential compared to planktonic bacteria and LPS. EPS produced higher levels of inflammatory response compared to eDNA for both strains (P < 0.05). IL-6 and TNF-α, and NO expression showed no difference for E. faecalis EPS (P ≥ 0.05). In contrast, P. aeruginosa EPS and eDNA had significant levels of IL-6 compared to TNF-α and NO (P < 0.05). CONCLUSIONS: Monospecies biofilm matrix EPS and eDNA from the bacterial strains tested had the ability to induce a low-grade inflammatory response when compared to planktonic bacteria and LPS. This study highlights the potential of biofilm matrix/components, devoid of bacteria to induce low-grade chronic inflammation.

Xenobiotic metabolizing enzyme activities in cells used for testing skin sensitization in vitro.

For ethical and regulatory reasons, in vitro tests for scoring potential toxicities of cosmetics are essential. A test strategy for investigating potential skin sensitization using two human keratinocytic and two human dendritic cell lines has been developed (Mehling et al. Arch Toxicol 86:1273­1295, 2012). Since prohaptens may be metabolically activated in the skin, information on xenobiotic metabolizing enzyme (XME) activities in these cell lines is of high interest. In this study, XME activity assays, monitoring metabolite or cofactor, showed the following: all three passages of keratinocytic (KeratinoSens® and LuSens) and dendritic (U937 und THP-1) cells displayed N-acetyltransferase 1 (NAT1) activities (about 6­60 nmol/min/mg S9-protein for acetylation of para-aminobenzoic acid). This is relevant since reactive species of many cosmetics are metabolically controlled by cutaneous NAT1. Esterase activities of about 1­4 nmol fluorescein diacetate/min/mg S9-protein were observed in all passages of investigated keratinocytic and about 1 nmol fluorescein diacetate/min/mg S9-protein in dendritic cell lines. This is also of practical relevance since many esters and amides are detoxified and others activated by cutaneous esterases. In both keratinocytic cell lines, activities of aldehyde dehydrogenase (ALDH) were observed (5­17 nmol product/min/mg cytosolic protein). ALDH is relevant for the detoxication of reactive aldehydes. Activities of several other XME were below detection, namely the investigated cytochrome P450-dependent alkylresorufin O-dealkylases 7-ethylresorufin O-deethylase, 7-benzylresorufin O-debenzylase and 7-pentylresorufin O-depentylase (while NADPH cytochrome c reductase activities were much above the limit of quantification), the flavin-containing monooxygenase, the alcohol dehydrogenase as well as the UDP glucuronosyl transferase activities.

Comparative study of BCR-ABL1 quantification: Xpert assay, a feasible solution to standardization concerns.

The level of BCR-ABL1 reached after treatment with tyrosine kinase inhibitors is an effective marker of the therapeutic response and a good survival predictor in chronic myeloid leukemia (CML) patients. However, no agreement has yet been achieved about either the standardization of the technique to determine BCR-ABL1 or the interpretation of the results. The aim of this study was to compare the method currently recommended by the European Leukemia Net, which includes the application of a conversion factor to express the results in international scale, with an automated method (Xpert BCR-ABL™, Cepheid). BCR-ABL1 transcript quantification was performed in 117 samples from CML patients in two different laboratories by both methods, and the results were compared by statistical procedures. A high linear correlation was obtained in the results between the two methods. The concordance at logarithmic intervals reached 62 %. When the major molecular response (MMR) was analyzed, 85 % agreement was achieved. The automated method provides reproducible results and does not show significant differences compared with the traditional method. As a clinical tool, Xpert correctly classified the patients in MMR and can be considered a useful alternative for the molecular follow-up of CML patients.

First Report of Dolabra nepheliae Associated with Corky Bark Disease of Rambutan and Pulasan in Honduras.

In the last decade, rambutan (Nephelium lappaceum L., Sapindaceae) and pulasan (N. mutabile Blume) have been cultivated in Honduras to produce exotic fruits for export to North America (2). Recently, a disease was observed that produces dark brown to black fissured cankers from 1 to 3 cm long and 1 to 4 cm wide. The infected bark tissue becomes swollen with the middle region 3 to 8 mm thick. Symptoms appear when the trees are approximately 3 years old. As the trees mature, the cankers increase in size and weaken the branches, often resulting in breakage with the weight of the fruit causing substantial plant damage and fruit loss. In August 2010, fissured branch samples of rambutan and pulasan were collected from 6- to 8-year-old trees from the Humid Tropical Demonstrative Agroforestry Center in Honduras, Atlantida, La Masica (15°33'47.4″N, 87°05'2.5″W, elevation 106 m). A fungus associated with the cankers was identified as Dolabra nepheliae. It produces black, stipitate, elongate ascomata, 312 to 482 × 250 to 281 µm with broadly cylindric, bitunicate asci, 120 to 138 × 11.2 to 15.0 µm, and filiform, hyaline ascospores, 128 to 135 × 2.8 to 3.2 µm. Fungi from rambutan and pulasan were isolated on cornmeal agar plus 0.5% dextrose and antibiotics. On potato dextrose agar, the ascospores produced slow-growing colonies, 5 mm per week. In culture, isolates from both hosts produced pycnidia with elongated, slightly to strongly curved or S-shaped, hyaline conidia, 22.8 to 46.4 × 2.8 to 3.7 µm. This fungus was first reported on rambutan and pulasan from Malaysia (1,4), and later reported on rambutan and litchi in Hawaii and Puerto Rico (3). To our knowledge, this is the first report of D. nepheliae on pulasan and rambutan from Honduras. Specimens have been deposited at the U.S. National Fungus Collections (BPI 882442 on N. lappaceum and BPI 882443 on N. mutabile). Cultures were deposited at the Centraalbureau voor Schimmelcultures (CBS) as CBS 131490 on N. lappaceum and CBS 131491 on N. mutabile. Sequences of the internal transcribed spacer (ITS) region including ITS1, 5.8S, and ITS2 intergenic spacers were deposited in GenBank (Accession No. JQ004281 on N. lappaceum and Accession No. JQ004280 on N. mutabile). A BLAST search and pairwise comparison using the GenBank web server were used to compare ITS sequence data and recovered the following results: (i) CBS 131490 on N. lappaceum is 99% (538 of 544) identical to D. nepheliae CBS 123297 on Litchi chinensis from Puerto Rico; and (ii) CBS 131491 on N. mutabile is 99% (527 of 533) identical to the same strain of D. nepheliae. On the basis of the ITS sequence data, the isolates from Honduras were confirmed as the same species, D. nepheliae from Puerto Rico. Efforts to develop resistant germplasm and management strategies to control this disease have been initiated. References: (1) C. Booth and W. P. Ting. Trans. Brit. Mycol. Soc. 47:235, 1964. (2) T. Ramírez et al. Manual Para el Cultivo de Rambutan en Honduras. Fundación Hondureña de Investigación Agrícola. La Lima, Cortes, Honduras, 2003. (3) A. Y. Rossman et al. Plant Dis. 91:1685, 2007. (4) H. Zalasky et al. Can. J. Bot. 49:559, 1971.

Experimental design for the optimization of copper biosorption from aqueous solution by Aspergillus terreus.

An experimental design methodology was applied to study the effects of temperature, pH, biomass dose, and stirring speed on copper removal from aqueous solutions by Aspergillus terreus in a biosorption batch system. To identify the effects of the main factors and their interactions on copper removal efficiency and to optimize the process, a full 2(4) factorial design with central points was performed. Four factors were studied at two levels, including stirring speed (50-150 min(-1)), temperature (30-50°C), pH (4-6) and biosorbent dose (0.01-0.175 g). The main factors observed were pH and biomass dose, along with the interactions between pH and biomass, and stirring speed. The optimal operational conditions were obtained using a response surface methodology. The adequacy of the proposed model at 99% confidence level was confirmed by its high adjusted linear coefficient of determination (R(Adj)(2)=0.9452). The best conditions for copper biosorption in the present study were: pH 6, biosorbent dose of 0.175 g, stirring speed of 50 min(-1) and temperature of 50°C. Under these conditions, the maximum predicted copper removal efficiency was 68.52% (adsorption capacity of 15.24 mg/g). The difference between the experimental and predicted copper removal efficiency at the optimal conditions was 4.8%, which implies that the model represented very well the experimental data.

Preliminary study of the inflammatory response to subcutaneous implantation of three root canal sealers.

AIM: To evaluate the kinetics of the inflammatory tissue response to three root canal sealers using a physicochemical method for quantification of the enhanced vascular permeability and histopathological analysis. METHODOLOGY: Twenty-eight male Wistar rats randomly assigned to four groups according to the evaluation periods (1, 3, 7 and 14 days) were used to assess the vascular permeability and histopathological reaction to RoekoSeal, AH Plus and Sealapex (new formulation) sealers, using saline and Chloropercha as negative and positive controls, respectively. Seven rats were sacrificed per period. The biocompatibility of the sealers was evaluated spectrophotometrically and histopathologically. RESULTS: At day 14, Sealapex produced significantly more inflammatory exudate than AH Plus and RoekoSeal (P < 0.05); however, there was no significant difference between AH Plus and RoekoSeal (P > 0.05). Sealapex (new formulation) was the most irritating sealer, producing severe inflammation with the presence of multinucleated giant cells. RoekoSeal was the most biocompatible sealer, producing the least amount of inflammatory exudate. CONCLUSIONS: RoekoSeal root canal sealer was biocompatible when implanted in connective tissue.

Brimonidine-LAPONITE® intravitreal formulation has an ocular hypotensive and neuroprotective effect throughout 6 months of follow-up in a glaucoma animal model.

Intravitreal administration is widely used in ophthalmological practice to maintain therapeutic drug levels near the neuroretina and because drug delivery systems are necessary to avoid reinjections and sight-threatening side effects. However, currently there is no intravitreal treatment for glaucoma. The brimonidine-LAPONITE® formulation was created with the aim of treating glaucoma for extended periods with a single intravitreal injection. Glaucoma was induced by producing ocular hypertension in two rat cohorts: [BRI-LAP] and [non-bri], with and without treatment, respectively. Eyes treated with brimonidine-LAPONITE® showed lower ocular pressure levels up to week 8 (p < 0.001), functional neuroprotection explored by scotopic and photopic negative response electroretinography (p = 0.042), and structural protection of the retina, retinal nerve fibre layer and ganglion cell layer (p = 0.038), especially on the superior-inferior axis explored by optical coherence tomography, which was corroborated by a higher retinal ganglion cell count (p = 0.040) using immunohistochemistry (Brn3a antibody) up to the end of the study (week 24). Furthermore, delayed neuroprotection was detected in the contralateral eye. Brimonidine was detected in treated rat eyes for up to 6 months. Brimonidine-LAPONITE® seems to be a potential sustained-delivery intravitreal drug for glaucoma treatment.

Peptide reactivity associated with skin sensitization: The QSAR Toolbox and TIMES compared to the DPRA.

The molecular initiating event (MIE) of skin sensitization is the binding of a hapten to dermal proteins. This can be assessed using the in chemico direct peptide reactivity assay (DPRA) or in silico tools such as the QSAR Toolbox and TIMES SS. In this study, the suitability of these methods was analyzed by comparing their results to in vivo sensitization data of LLNA and human studies. Compared to human data, 84% of non-sensitizers and sensitizers yielded consistent results in the DPRA. In silico tools resulted in 'no alert' for 83%-100% of the non-sensitizers, but alerted only 55%-61% of the sensitizers. The inclusion of biotic and abiotic transformation simulations yielded more alerts for sensitizers, but simultaneously dropped the number of non-alerted non-sensitizers. In contrast to the DPRA, in silico tools were more consistent with results of the LLNA than human data. Interestingly, the new "DPRA profilers" (QSAR Toolbox) provided unsatisfactory results. Additionally, the results were combined in the '2 out of 3' prediction model with in vitro data derived from LuSens and h-CLAT. Using DPRA results, the model identified 90% of human sensitizers and non-sensitizers; using in silico results (including abiotic and biotic activations) instead of DPRA results led to a comparable high predictivity.

Importance of salt fingering for new nitrogen supply in the oligotrophic ocean.

The input of new nitrogen into the euphotic zone constrains the export of organic carbon to the deep ocean and thereby the biologically mediated long-term CO2 exchange between the ocean and atmosphere. In low-latitude open-ocean regions, turbulence-driven nitrate diffusion from the ocean's interior and biological fixation of atmospheric N2 are the main sources of new nitrogen for phytoplankton productivity. With measurements across the tropical and subtropical Atlantic, Pacific and Indian oceans, we show that nitrate diffusion (171±190 µmol m(-2) d(-1)) dominates over N2 fixation (9.0±9.4 µmol m(-2) d(-1)) at the time of sampling. Nitrate diffusion mediated by salt fingers is responsible for ca. 20% of the new nitrogen supply in several provinces of the Atlantic and Indian Oceans. Our results indicate that salt finger diffusion should be considered in present and future ocean nitrogen budgets, as it could supply globally 0.23-1.00 Tmol N yr(-1) to the euphotic zone.

Possible association between Taenia solium cysticercosis and cancer: increased frequency of DNA damage in peripheral lymphocytes from neurocysticercosis patients.

Helminths, particularly some Schistosoma species, have been associated with cancer in humans. Neurocysticercosis, produced by cysticerci of the helminth Taenia solium, has been associated with the emergence of brain tumours and haematological malignancies. Local tumours, such as glioblastoma, could be explained by the induction of DNA damage in cells surrounding the cysticercus and chronically exposed to an inflammatory host response. However, systemic effects such as haematological malignancies are not easy to understand. The present work was conducted in Mexico to find out whether DNA damage arises in peripheral lymphocytes in patients with neurocysticercosis. We utilized a highly sensitive technique to analyse chromosomal aberrations, in-situ hybridization with probes against chromosomes 1, 2 and 4, and in addition the blocked-cytokinesis technique was used to determine the formation of micronuclei, a peculiar form of DNA damage. The study was made in lymphocytes from 8 patients before and after the administration of praziquantel, 1 of the 2 drugs used for neurocysticercosis treatment. The frequencies of chromosome aberrations and micronuclei in peripheral blood lymphocytes were higher in the infected patients as compared to those observed both in healthy donors and in the group of patients after praziquantel therapy. Our results suggest that chromosome aberrations induced in peripheral cells during neurocysticercosis could be associated with the development of haematological neoplasias.

Pathways and roadblocks in muscarinic receptor-mediated growth regulation.

In some cell systems muscarinic receptor stimulation can induce proliferation or transformation. This phenomenon is subtype-specific (only m1 and m3 receptors are effective) and cell type dependent. In 1321N1 astrocytoma cells activation of m3 receptors stimulates phospholipase C, but does not induce DNA synthesis. In contrast the thrombin receptor, which also couples to phospholipase C, is strongly mitogenic and induces AP-1-dependent gene expression. Various experimental findings indicate that this discrepancy is not due to muscarinic receptor desensitization or blockade of growth stimulatory pathways. Muscarinic receptor number may be limiting, in particular for receptor coupling to the pertussis toxin-insensitive G-protein G12. This G-protein is required for thrombin-induced mitogenesis in 1321N1 cells and may couple selectively to the thrombin versus muscarinic receptor. In cardiomyocytes hypertrophic cell growth is induced by heterologously expressed m1 or m3 receptors but not by the endogenous m2 receptors. Studies using chimeric receptors confirm that induction of hypertrophy requires signalling through phospholipase C, but indicate that additional signals are needed to induce the morphological features of this response. We suggest that small G-proteins of the Rho subfamily, in addition to G12, mediate growth responses to G-protein-coupled receptors.

Ca 15.3 in breast cancer: comparison of two assays and validation in a Mexican population.

Ca 15.3 is a tumor marker used for breast carcinoma, since one epitope is an antigen present in milk fat globules. Serum from 171 patients with breast cancer upon initial presentation was studied for Ca 15.3. In the first 72 cases, the authors compared RIA vs. ELISA using a simple linear regression. On the following 99, only ELISA was performed. With all 171 patients, a clinical association between Ca 15.3 measurement and age, stage and hormone receptors was carried out. Correlation coefficient between RIA and ELISA was 0.85. Of 104 patients below 50 years of age, 88 had normal Ca 15.3 and 16, elevated; 67 were older than 50 years, 46 had normal Ca 15.3 and 21, elevated (p=0.022). Ca 15.3 was elevated in 11% of patients with clinical stages I/II, and 89% in stages III/IV (p=0.0001). The association of Ca 15.3 with hormone receptors was not significant. In conclusion, ELISA and RIA measure Ca 15.3 with comparable results, the first method has the advantage of not using radioactivity. The authors found higher probability of elevated Ca 15.3 in older patients and in those with advanced disease.

In vitro effects of albendazole and its metabolites on the cell proliferation kinetics and micronuclei frequency of stimulated human lymphocytes.

BACKGROUND: Albendazole (ABZ) is an antiparasitic drug used for the treatment of several helminthiases. After its oral administration, this compound is metabolized to sulfoxide (SOABZ) and sulfone (SO(2)ABZ), SOABZ being the active metabolite. The antiparasitic activity of ABZ has been associated with its capacity to bind with tubulin, altering microtubule formation. Although some studies indicate that ABZ modified microtubule structure in host cells, data concerning the consequences of this phenomenon in human cells are scant. METHODS: In this study we evaluated the effects of ABZ and its metabolites on cell proliferation, as well as on the frequency of micronucleated cells in cultured human lymphocytes. RESULTS: ABZ and SOABZ arrested cell proliferation in metaphase and increased the frequency of micronuclei in treated lymphocytes. Contrariwise, SO(2)ABZ, the inactive metabolite, did not produce any significant effect. CONCLUSIONS: The formation of micronuclei may ultimately result in aneuploidy induction, an effect that could have severe consequences in humans. However, the doses of ABZ and SOABZ at which these effects were observed are several orders of magnitude higher than those found in the plasma of treated individuals. Because there are other mechanisms by which aneuploidy can be induced at even lower doses than micronuclei, i.e., chromosome nondisjunction, it is necessary to evaluate this effect in exposed individuals.

The ambrosanolide cumanin inhibits macrophage nitric oxide synthesis: some structural considerations.

Since its role in inflammatory diseases was recognized, nitric oxide (NO) has become an important mediator to evaluate anti-inflammatory agents. Sesquiterpene lactones, which occur in several medicinal plants, inhibit the NO production in macrophage-like cells. This action is probably due to a 1,4 addition reaction between its alpha,beta-unsaturated carbonyl group with sulfhydryl (SH)-containing compounds. For this reason it is believed that these compounds are cytotoxic, which restricts their therapeutic use. In this contribution, the ability of the ambrosanolide-type sesquiterpene lactone cumanin (from the Asteraceae Ambrosia psilostachya) to inhibit NO biosynthesis was evaluated in lipopolisaccharide-induced peritoneal murine macrophages and its cytotoxicity was assessed in the MTT assay. Cumanin showed a potent inhibitory effect in NO production (IC(50) = 9.38+/-0.38 microM) with low cytotoxicity. The 1,4-addition reaction of thiols was slow, which does not explain the inhibition of NO production but does explain the low cytotoxicity of cumanin with respect to other lactones.

Coley toxins immunotherapy: a retrospective review.

OBJECTIVE: Coley toxins are administered to cancer patients worldwide, though clinical studies assessing efficacy either alone or in combination with conventional cancer therapy are limited. This article provides an overview of Coley toxins immunotherapy and compares the survival experience of cancer patients who received Coley toxins for renal, ovarian, breast cancer, or soft-tissue sarcomas with patients who received conventional treatment other than radiation. DATA SOURCES: Cases were compiled from 5 of 18 monographs by Helen Coley Nauts. STUDY SELECTION: Using a retrospective cohort design with external controls, 128 Coley cases treated in New York from 1890 to 1960 were compared with 1675 controls from the Surveillance Epidemiology End Result (SEER) population-based cancer registry who received a cancer diagnosis in 1983. DATA EXTRACTION: Groups were matched on age, sex, ethnicity, site, stage, and treatment status (i.e., no radiotherapy). DATA SYNTHESIS: The Cox proportional hazards model controlled for stage and menopausal status (when applicable) and the hazard ratio and 95% CI defined the odds of site-specific survival from date of diagnosis to last follow-up. Compared to the SEER population, risk of death within 10 years was not significantly different in Coley patients treated for renal, ovarian, breast cancer, or soft-tissue sarcomas. CONCLUSIONS: This study suggests that patients treated with surgery and Coley toxins between 1890 and 1960 experienced survival rates comparable to those of patients diagnosed in 1983 and treated with nonradiotherapeutic conventional approaches. The study is limited by small sample sizes, possibly inaccurate technology for staging during Coley time, and potential selection bias with Coley patients.

Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease): diagnosis with fine-needle aspiration in a case with nodal and nasal involvement.

Sinus histiocytosis with massive lymphadenopathy (SHML) is a rare entity characterized by the proliferation of cells belonging to the macrophage-histiocyte family, but whose exact origin is unknown. A case of SHML occurring in a 62-yr-old female is described in which the diagnosis is initially suggested by fine-needle aspiration (FNA) of a cervical node. The smears show a background of lymphocytes and plasma cells and large histiocytes with well-preserved lymphocytes in their cytoplasm (emperipolesis or lymphophagocytosis). Both nodal and extranodal (nasal mass) involvement are confirmed by surgical biopsy. The immunohistochemistry suggests that SHML cells are functionally activated macrophages. Eight months after diagnosis, the patient is clinically well, with partial improvement of lymphadenopathy.

Granular dystrophy associated with congenital cataracts.

Seven members of the same family suffering from Groenow I granular dystrophy and associated congenital cataracts have been studied. Three patients had corneal dystrophy at various stages and three others had a fetal nuclear congenital cataract. A penetrating keratoplasty button was studied under light and electron microscopies. This association has not been described previously. However we must emphasize that there is no general validity for a correlation between granular dystrophy and congenital cataract.

Morphometric study of wound healing in a model of filtering surgery with mitomycin-C.

BACKGROUND: Failure of filtering surgery may be related to excessive wound healing in the surgical area. This effect diminishes with the use of antimetabolic agents. Mitomycin-C (MMC) has proved to be the most effective drug to reduce myofibroblastic proliferation in experimental in vivo and in vitro models. To our knowledge, the objective changes induced by mitomycin-C in the size of wound healing areas have not been investigated. METHODS: Filtering surgery was performed on both eyes of 40 pigmented rabbits. Preoperatively one of the eyes received MMC (0.5 mg/ml), and the fellow eye received balance salt solution as placebo. Animals were killed on days 6, 15, 30 and 58. Microscopic healing areas were measured by digital procedures. The areas of target and fellow control eyes were compared by the Wilcoxon test. RESULTS: This study showed significant differences (p < 0.05) between treated and untreated groups, the healing are a gradually becoming smaller. CONCLUSIONS: Objective methods to quantify the microscopic effects of MMC can be useful to improve our knowledge about the action on this antimetabolite and to enable us to adjust more accurately the timing and dosage when applying the drug in glaucoma filtering surgery.

Long-term effects of postoperative subconjunctival injections of mitomycin-C in the rabbit eye.

AIM: Single or multiple subconjunctival injections of mitomycin-C (MMC) may offer one way of establishing the total dosage of MMC more accurately. The method also allows re-applications later postoperatively. In this experimental, randomized prospective study we compared the effects of a single intraoperative application of MMC at the filtering site and a single postoperative subconjunctival injection of the drug. METHODS: The left eyes of 32 pigmented rabbits were divided into two groups. In the first group we applied MMC intraoperatively (IO) with a 4 x 1 mm surgical sponge soaked in a MMC solution (0.5 mg/ml). In the second group we injected 0.4 ml of the same solution subconjunctivally (SC) immediately after (conjunctival) suture. Post-operative evaluation was carried out every day during the first week, then every three days until day 58. Survival analyses were done for intraocular pressure (IOP) and bleb failure. Log-rank tests were used to compare survival differences between the groups. RESULTS: The IO group showed longer survival parameters than the SC group (p < 0.05), both in the control of IOP and as regards blebs. The histological persistence of fistulas was similar. The IO group, however, had a higher incidence of undesirable side effects. CONCLUSIONS: Our findings suggest IO application of MMC is more effective in reducing fibroblast ingrowth. However, subconjunctival application offers certain advantages such as the possibility of repeating the treatment postoperatively and, therefore, using a smaller initial dose.

Study of Medicare beneficiary complaint procedures.

New procedures for reviewing a sample of Medicare beneficiary complaints about quality of care are compared with traditional procedures at a peer review organization (PRO) for 1998-1999. These new procedures included: (1) expanded communications with complainants and providers, (2) changed data collection methods, (3) integrated concurrent review findings from other agencies, (4) expedited review procedures, and (5) changed the medical review procedures. The findings showed improved beneficiary satisfaction with the new procedures over the traditional procedures and shorter time periods for processing the reviews. Even with the new procedures, beneficiaries continued to be concerned that the review time frames were too lengthy, the reviews generally failed to confirm their complaints, and the PROs generally did not disclose the findings to the beneficiaries.