Potential chitosan-coated alginate nanoparticles for ocular delivery of daptomycin.

Daptomycin may offer an antibacterial alternative for the treatment of endophthalmitis caused by methicillin-resistant Staphylococcus aureus (MRSA) and other potential agents. In the present project, mucoadhesive chitosan-coated alginate (CS-ALG) nanoparticles are proposed as an effective delivery system for daptomycin permeation across ocular epithelia, with potential for the treatment of bacterial endophthalmitis. CS-ALG nanoparticles were prepared by ionotropic pre-gelation of an alginate core followed by chitosan polyelectrolyte complexation, and characterized regarding particle size, polydispersity, and zeta potential. The encapsulation efficiency was determined and antimicrobial activity was also tested after encapsulation of the antibiotic. Also, in vitro ocular permeability of free daptomycin and encapsulation into chitosan and CS-ALG nanoparticles was evaluated using ocular epithelial cell culture models. Formulated daptomycin-loaded CS-ALG nanoparticles were negatively charged, with a size range of 380-420 nm, suitable for ocular application. The encapsulation efficiency was between 79 and 92 %, with decreasing alginate:daptomycin mass ratios. The antibacterial activity of daptomycin against major microorganisms responsible for bacterial endophthalmitis was not affected by encapsulation into nanoparticles. Daptomycin permeability was up to 16 % (chitosan nanoparticles) and 9 % (CS-ALG nanoparticles) through corneal cell monolayer, and 18 % (chitosan nanoparticles) and 12 % (CS-ALG nanoparticles) for retinal cell monolayer after 4 h, demonstrating epithelial retention of the drug compared to free drug. The developed daptomycin-loaded CS-ALG nanoparticles seem to be an interesting and potential system for ocular daptomycin delivery and treatment of bacterial endophthalmitis.

Cold modalities with different thermodynamic properties have similar effects on muscular performance and activation.

Although tissue cooling is widely used in the treatment of musculoskeletal injuries there is still controversy about its effects on muscular performance. The combination of cooling and exercise justifies the study of this topic. The aim was to compare the effects of ice pack and cold-water immersion on the muscular performance parameters of plantar flexors and muscular activation of the triceps surae. 41 healthy men (mean age: 22.1 years, SD: 2.9) were randomly assigned to cooling with either ice pack (n=20) or cold-water immersion (n=21). Independent variables were cold modality (ice pack or cold-water immersion) and pre- and post-cooling measurement time. Dependent variables were muscular performance (measured during isometric and concentric contractions of plantar flexors) and electromyography parameters of the triceps surae (median frequency and root mean square amplitude). Dependent-samples t-tests were used to compare pre- and post-cooling data and independent-samples t-tests were used to compare the difference (pre- and post-cooling) between groups. Ice pack increased isometric peak torque (mean: 9.00 Nm, P=0.01) and both cold modalities reduced muscular activation in triceps surae (P<0.0001); Cold-water immersion and ice pack reduced peak torque and total work during dynamic isokinetic contraction at both velocities (mean: -11,00 Nm, P<0.05) and affected muscular activation in different ways. In conclusion, ice pack increases isometric torque, while both ice pack and cold-water immersion decrease concentric muscular performance. These results indicate that these cooling methods should be chosen with caution, considering the type of task required during training or rehabilitation. New studies investigating other muscle groups and joints are necessary.

Different cellular and immunohistochemical abdominal wall cicatrization parameters evaluation in comparison with sublay, onlay, and ipom technique in an experimental rat model.

PURPOSE: Incisional hernia (IH) occurs when there is a partial or complete solution of continuity of a fascia previously incised. Systematic reviews demonstrate that surgical treatment of IHs with the use of meshes are approximately 16%. Meta-analyses have demonstrated the superiority of mesh placement using sublay technique, but without a pathophysiological explanation. Thus, we aim to evaluate the different techniques of mesh positioning in an experimental model. METHODS: Fifty rats were distributed into five groups; control; simulation (SM)-submitted to laparotomy only; onlay-the mesh was positioned in onlay fashion; retromuscular (SL)-the mesh was positioned in a sublay fashion; intraperitoneal (IPOM)-positioning of the mesh adjacent to the transversalis fascia, inside the cavity. After 60 days, adhesions, tensiometry, histology, and immunohistochemistry were addressed. RESULTS: The IPOM group had the most adhesions, together with the SL group, with significantly relevant results. The SL group had higher values of tensiometric evaluation, while the IPOM group had the lowest mean in the tensiometry evaluation, being even lower than the SM group. Regarding histological and immunohistochemical findings, the SL group had a higher pixel number count compared to the groups, with statistical significance, in addition to higher expression of polymorphonuclear infiltrate and CD68 markers. CONCLUSION: The mesh positioning in sublay compartment is associated with the development of more pronounce minimum tensile force required for detaching the surrounding abdominal wall tissues it was incorporated. The intensity of these findings correlates to the different histological and immunohistochemical profiles observed following each repair, since SL group was characterized by a higher proportion of collagen, inflammatory, and reparative elements. Characterizing these pro-healing elements and its counterparts will allow the development of new therapeutic tools which could be added to the still far-from-ideal current therapeutic options for IH treatment.

Simultaneous detection and identification of the Xanthomonas species complex associated with tomato bacterial spot using species-specific primers and multiplex PCR.

AIMS: To establish protocols for the simultaneous detection and identification of Xanthomonas species causing tomato bacterial spot. METHODS AND RESULTS: We verified the specificity and sensitivity of the previously reported sets of primers designed for strains of the four species of Brazilian tomato bacterial spot xanthomonads, consisting of 30 of Xanthomonas euvesicatoria, 30 of X. vesicatoria, 50 of X. perforans and 50 of X. gardneri. Furthermore, we tested a multiplex PCR protocol for the purpose of concurrent species identification. The possibility of direct detection of the pathogens in diseased leaf samples was also verified. The primers were highly specific, amplifying only target DNA. The sensitivity of the primers in conventional PCR was 50 pg µl(-1) for purified DNA and ranged from 5 × 10(2) to 5 × 10(4) CFU ml(-1) when bacterial suspensions were analysed. The multiplex PCR was suitable for the detection of all four species and showed similar sensitivity to conventional PCR when tested on purified DNA. When using bacterial suspensions, its sensitivity was similar to conventional PCR only when a biological amplification step (Bio-PCR) was included. Both methods were able to detect the pathogens in symptomatic tomato leaves. CONCLUSIONS: Brazilian Xanthomonas strains causing tomato bacterial spot can be differentiated and identified at species level by a PCR-based method and by a multiplex PCR. SIGNIFICANCE AND IMPACT OF THE STUDY: This protocol may be a feasible alternative tool for the identification and detection of these pathogens in plant material and may be used for routine diagnostic purposes in plant pathology laboratories.

External auditory canal exostoses: long-term surgical satisfaction and its relationship with surgical complications.

OBJECTIVE: The main purpose of the present study was to evaluate whether complications related to surgery for exostoses are associated with a decrease in patients' quality of life. METHODS: This was a retrospective study for which the following information was collected: sex, age, pre- and post-operative symptoms, pre- and post-operative audiological evaluation results, surgical approach, instruments used, complications, and Glasgow Benefit Inventory score. RESULTS: The study included 67 patients (94 ears). The three main complaints reported were wax retention, otitis externa and hearing loss. Surgical complications occurred in 14.9 per cent of patients. Patients experienced a significant benefit from surgery, especially in relation to somatic state, with a global Glasgow Benefit Inventory score of + 44.3. No significant difference was found between the global Glasgow Benefit Inventory changes and surgery-related complications (p = 0.093). CONCLUSION: After surgery for exostoses, the vast majority of patients showed improvement. Complications related to surgery in general do not seem to influence patients' satisfaction with surgery.

Genotoxic evaluation of different doses of inorganic lead (PbII) in Hoplias malabaricus.

Different genetic biomarkers have been used to evaluate the pollution effects of mutagenic agents such as metals and also a great variety of chemicals delivered on the environment by human activities. This way, the aim of the present report was to evaluate the effects of inorganic lead in fishes through the frequency of piscine micronuclei and nuclear morphological alterations in peripheral cells, chromosomal aberration frequency and comet assays in blood and kidney cells. Specimens of Hoplias malabaricus received different doses of lead by intra-peritoneal injections at time of 96 h. There was not a significant difference between control and treated groups for the piscine micronucleus and chromosomal aberration assays. In the comet assays there was a significant difference between control and contaminated groups. However, a significant difference between the applied doses was not observed. The results obtained with the comet assays also show that blood presented a higher sensibility than the kidney tissue, possibly due to the acute contamination. Although the results showed the genotoxic potential of lead at the 21 and 63 microg Pb(2 + )/g doses for both tissues, the lowest dose is considered more appropriate for future bioassays.

Radiological evaluation of facial types in mouth breathing children: a retrospective study.

Mouth breathing is a condition often associated with a long face, half-open mouth and increased anterior facial height. We performed conventional lateral and frontal cephalograms of eighty-nine children with nasal and mouth breathing and independently measured Total Facial Height using the analysis technique of Ricketts, and the Morphologic Facial Index employing the technique of Avila. It was concluded that dolicofacial following mesofacial were the most frequent patterns found in mouth-breathing children and this suggests that both analyses can be used independently.

Use of hepatocytes from Hoplias malabaricus to characterize the toxicity of a complex mixture of lipophilic halogenated compounds.

Organisms are continuously exposed to a plethora of anthropogenic toxicants daily released to the environment. In the present study, the effects of a mixture of halogenated organic compounds (HOCs) extracted from hepatic lipids were evaluated on the primary hepatocyte culture from fish Hoplias malabaricus. Cells were isolated through non-enzymatic perfusion protocol and cultured during 3 days to allow attachment. Two concentrations of the mixture of HOCs (10 ng ml(-1) [Mix10] and 50 ng ml(-1) [Mix50]) were tested in cells for 2 days by medium replacement. The control groups, with and without solvent (DMSO) were run in the same conditions. Both tested concentrations of HOCs increased the catalase and GST activities, but only the Mix50 increase the DNA damage and decreased the GSH concentration and cell viability. Lipid peroxidation increased in the Mix10 group, but it seems to be more a consequence of DMSO presence than the HOCs themselves. The DMSO at 0.1% increased the lipid peroxidation, GSH concentration, apoptosis and DNA damage. The present data suggest that DMSO interferes with the hepatocytes of H. malabaricus in culture and that the mixture of HOCs tested alters the redox state of the hepatocytes.

Acute renal failure in cirrhosis associated with macroscopic hematuria of glomerular origin.

Two patients with alcoholic cirrhosis developed an acute impairment of renal function during an episode of macroscopic hematuria, one of them requiring hemodialysis treatment. Later, when gross hematuria disappeared, renal function gradually improved. The histological findings in both patients showed a mild mesangial glomerulonephritis with IgA deposits on immunofluorescence, the presence of red blood cells casts obstructing the lumen in 40% to 50% of the renal tubules, and a marked tubular necrosis.

Relapse of tuberculosis after treatment in human immunodeficiency virus-infected patients.

OBJECTIVES: To evaluate the relapse rate of tuberculosis after a complete course of antituberculous therapy in human immunodeficiency virus-infected patients and to identify the risk factors for relapse. PATIENTS AND METHODS: Historic cohort study of all adult patients who were diagnosed as having human immunodeficiency virus infection and a first episode of culture-proved tuberculosis at 2 university hospitals in Madrid, Spain, between 1986 and 1992, and who completed at least 6 months of treatment were included and followed up until September 1994. RESULTS: Of 276 patients with human immunodeficiency virus infection and tuberculosis, 87 could not be evaluated (6 died before treatment, 39 died during treatment, 36 did not complete the planned therapy, and 6 were unavailable during treatment). The remaining 189 received a standard regimen (ie, 3 or 4 drugs, always including rifampin and isoniazid, for > or = 6 months). The median duration of follow-up for these 189 patients was 31.5 months, with a total of 4668 patient-months of follow-up after treatment; 105 patients (56%) were followed up until death. The relapse rate was 7.9% (2.7/100 patient-years). With multivariate analysis, a shorter duration of treatment and a low CD4+ cell count were associated with a greater probability of relapse. Relapses occurred in 5 (3.4%) of 148 patients who were treated for 9 or more months (1.7/100 patient-years) and in 10 (24%) of 41 patients who were treated for less than 9 months (10.9/100 patient-years) (P < .001; relative hazard, 9.2; 95% confidence interval, 3.1-26.9). CONCLUSIONS: As standard antituberculous therapy for 9 months is associated with a low rate of relapse, maintenance therapy is not required. Duration of treatment for less than 9 months is associated with a high rate of relapse.

The influence of hepatitis C virus-human immunodeficiency virus co-infection on the appearance of liver enzyme elevation in people on high activity antiretroviral treatment.

BACKGROUND: Liver enzyme elevation (LEE) as a consequence of HAART is a problem among patients with HIV-HCV co-infection. METHODS: In this retrospective study, 145 patients with HIV who were on HAART and who developed LEE grades 3 and 4 of the World Health Organization (WHO) were followed up. Basal ALT, alcohol consumption, and HCV and HBV co-infection were recorded. Comparisons were made between patients with and without HCV co-infection. RESULTS: Three patients without co-infection presented LEE grade 3 versus 38 with co-infection (104 episodes). An increase in basal ALT (RR: 1.01) and HCV co-infection (RR: 6.6) were the variables associated with LEE grade 3. The number of days that HAART had to be withdrawn due to LEE was 58.15 and 4.85 in subjects with and without co-infection, respectively (p=0.024). CONCLUSION: Patients with HCV-HIV co-infection have more episodes of LEE and must go longer without HAART than people without co-infection.

Mycobacterium fortuitum complex endocarditis-case report and literature review.

Endocarditis due to Mycobacterium fortuitum complex is a rare entity generally linked to the hospital environment. Only 18 cases have been published since 1966. Here we present a case of a female who developed an endocarditis due to Mycobacterium chelonae after valve replacement as well as a review of the literature. The course of this kind of endocarditis is generally subacute and the outcome is usually fatal. Blood cultures were positive in 75% of cases of metallic valve endocarditis, versus 20% in bioprostheses. The treatment must include antibiotics that have shown activity against these mycobacteria, such as amikacin, imipenem, cefoxitin, fluorinated quinolones and macrolides (especially clarithromycin). Surgical removal is recommended. Although the prognosis for the patient is poor, we should expect better outcomes with the use of new antibiotic regimens.

Paradoxical responses in a cohort of HIV-1-infected patients with mycobacterial disease.

SETTING: Paradoxical worsening or relapse of opportunistic infections has been described after initiation of highly active anti-retroviral therapy (HAART) in human immunodeficiency virus (HIV) infected patients. DESIGN: Retrospective study of a group of 33 HIV-infected patients with mycobacterial disease analysing the incidence and characteristics of patients with and without paradoxical response after starting HAART and/or mycobacterial treatment. RESULTS: Nine patients in the group had paradoxical response. No significant difference of baseline characteristics was observed in these patients. The decrease in viral load was significantly greater among patients with paradoxical response than in patients without. CONCLUSION: No clinical difference was found in the evolution of HIV-infected patients with mycobacterial disease after the resolution of the episode of paradoxical response.

Central retinal vein occlusion in a patient with thrombotic thrombocytopenic purpura.

PURPOSE: To describe the occurrence of central retinal vein occlusion in a patient with thrombotic thrombocytopenic purpura. METHODS: Case report. RESULTS: A 52-year-old woman developed central retinal vein occlusion in the acute phase of thrombotic thrombocytopenic purpura, followed by iris neovascularization only 1 month after the onset of central retinal vein occlusion. Color Doppler imaging of the affected eye showed an increased resistive index of the central retinal artery and reduced blood flow velocity of the central retinal vein compared to the fellow eye. CONCLUSION: We describe a rare association between central retinal vein occlusion and thrombotic thrombocytopenic purpura with rapid evolution of iris neovascularization. Our case demonstrates that close ophthalmologic attention is important in thrombotic thrombocytopenic purpura.

Cutaneous lesions in patients with visceral leishmaniasis and HIV infection.

Cutaneous lesions attributed to Leishmania are very seldom observed in classic Kala-Azar, but recently some reports have mentioned them in patients with HIV infection. We found cutaneous lesions whose biopsy disclosed the presence of Leishmania organisms in six patients of a group of 32 HIV patients with visceral Leishmaniasis. These lesions did not present a uniform or specific appearance, even though they tended to localize symmetrically on acral zones. They consisted of erythematous papules and hypopigmented macules on the dorsa of the hands, feet, and elbows; small subcutaneous nodules on the thighs; and erythematoviolaceous, scaly plaques on the face. These lesions accompanied in every case the other symptoms and/or signs of visceral leishmaniasis, responded to anti-leishmanial treatment, and were sometimes the first indicator of recurrence. The histopathological study was non-specific, but showed in every case the presence of abundant amastigotes within the dermal histiocytes and free in the dermis or subcutaneous tissue. Data from literature review are similar to ours.

Effect of hypervitaminosis A on the Harderian gland in rats. A morphologic and morphometric study.

Daily administration of 100 I.U. vitamin A/gram body weight to rats, for a period of 15 days, had the following effects: Body weight of rats treated with excess vitamin A was lower (84.0 g) than that of the controls (147.0 g). The weight of the Harderian gland was lower of rats injected with vitamin A (54.0 mg) than for the controls (87.0 mg). Histological examination of the Harderian gland in the hypervitaminotic animal showed smaller acini, lower cells, wide lumina and small nuclei when compared with the controls. These data were confirmed with morphometric techniques.

Effect of hypervitaminosis A on tongue muscles of the rat. A morphometric study.

Daily administration of 200 I.U. vitamin A/gram body weight to rats, for a period of 10 days, causes generalized hypertrophy of the tongue muscles. The data were confirmed with morphometric techniques, consisting of increased cytoplasm and decreased nuclei and empty spaces, obtained by the Chalkley technique, nuclei of similar volume, obtained with the use of karyometry, and larger areas of cross-sections of muscle fibers.

[Influence of a day care hospital on the incidence of hospital admission of AIDS patients]. / Influencia del hospital de día en los requerimientos de ingreso hospitalario de los pacientes con SIDA.

BACKGROUND: Day care units have become an usual way of medical care for AIDS patients. However, their influence on the incidence of hospital admissions has not been evaluated. METHODS: Observational and longitudinal study of a cohort of 308 patients with aids diagnosed between 1990 and 1994 and followed-up to June 1996. The incidence of hospital admissions according to the hospital of follow-up (with or without day care unit) was analyzed. A multivariate analysis of the number of hospital admissions was performed using regression model adjusted to a distribution of Poisson. RESULTS: After AIDS diagnosis, the incidence of hospital admissions was 108 per 100 patient-years of follow up (21 days as inpatient per patient-year). Those patients controlled in the hospital with day care unit have less hospital admissions (relative risk after adjusting by CD4+ cells count and type of diagnostic disease: 0.64; CI95% 0.55-0.76), and less days as inpatient through their follow-up (11 to 31 days less). There was no difference in survival among patients followed in both hospitals. CONCLUSIONS: A day care unit decrease the incidence of hospital admissions in aids patients. This positive impact is more evident in patients with lesser CD4+ cell counts.