Outbreak of follicular conjunctivitis associated with the diagnosis of anaplasmosis in a sheep herd.

OBJECTIVE: To report an outbreak of follicular conjunctivitis in a group of sheep diagnosed with Anaplasma spp., without any other co-infection. ANIMALS STUDIED: In all, 18 animals from a sheep head, males and females, from eight months to four years of age, were assessed for follicular conjunctivitis. PROCEDURES: The procedures performed included general physical and ophthalmological examinations; PCR evaluation for infectious agents; analysis of hematological parameters, microbiological tests of ocular swabs, coproparasitological examination, histopathological examination of conjunctival biopsy. RESULTS: All 18 animals had uni- or bilateral follicular conjunctivitis, and one animal also had unilateral uveitis. The results of microbiological analyzes were negative for Moraxella spp., Staphylococcus spp., and Pseudomonas spp., and PCR analysis results were negative for Chlamydia spp., Mycoplasma spp., and Toxoplasma gondii. Anemia, thrombocytopenia, lymphocytosis, and an inclusion body in some erythrocytes, compatible with Anaplasma and PCR analysis for Anaplasma spp. were positive. CONCLUSION: Anaplasmosis may be associated with follicular conjunctivitis in sheep and should be included in the differential diagnosis list and investigated in cases of conjunctivitis in herds.

Nanostructured lipid carriers loaded with an association of minoxidil and latanoprost for targeted topical therapy of alopecia.

Alopecia is a condition associated with different etiologies, ranging from hormonal changes to chemotherapy, that affects over 80 million people in the USA. Nevertheless, there are currently few FDA-approved drugs for topical treatment, and existing formulations still present skin irritation issues, compromising treatment adherence. This work aimed to develop a safe formulation based on nanostructured lipid carriers (NLC) that entrap an association of minoxidil and latanoprost and target drug delivery to the hair follicles. To do so, thermal techniques combined with FTIR were used to assess the chemical compatibility of the proposed drug association. Then, NLC with 393.5 ± 36.0 nm (PdI < 0.4) and +22.5 ± 0.2 mV zeta potential were produced and shown to entrap 86.9% of minoxidil and 99.9% of latanoprost efficiently. In vitro, the free drug combination was indicated to exert positive effects over human primary epidermal keratinocytes, supporting cell proliferation, migration and inducing the mRNA expression of MKI67 proliferation marker and VEGF - a possible effector for minoxidil-mediated hair growth. Interestingly, such a favorable drug combination profile was optimized when delivered using our NLC. Furthermore, according to the HET-CAM and reconstructed human epidermis assays, the nanoformulation was well tolerated. Finally, drug penetration was evaluated in vitro using porcine skin. Such experiments indicated that the NLC could be deposited preferentially into the hair follicles, causing a considerable increase in the penetration of the two drugs in such structures, compared to the control (composed of the free compounds) and generating a target-effect of approximately 50% for both drugs. In summary, present results suggest that hair follicle-targeted delivery of the minoxidil and latanoprost combination is a promising alternative to treat alopecia.

Latanoprost Loaded in Polymeric Nanocapsules for Effective Topical Treatment of Alopecia.

Latanoprost has recently been used to treat alopecia as it causes an increase in the capillary density of patients. This work presents for the first time the development of polymeric nanocapsules containing latanoprost for the topical treatment of alopecia. Poly-ε-caprolactone nanocapsules loading latanoprost were developed by nanoprecipitation of the polymer on the surface of drug oily nanodroplets. The method encapsulated 93.9 ± 0.4% of the drug into nanocapsules of 197.8 (± 1.2) nm (PdI = 0.15 ± 0.01). The nanosystem presented a zeta potential equal to - 30.1 ± 1.8 mV and was stable for at least 90 days when stored at 6°C. The colloidal aqueous dispersion was non-irritating, according to the in vitro HET-CAM test. The nanocapsules improved latanoprost accumulation into the hair follicles when topically applied on porcine skin, delivering 30% more drug to these skin structures relative to the control solution (P < 0.05). Also, with a simple manual massage, latanoprost accumulation was increased by twofold (P < 0.05). In conclusion, in addition to being a stable and safe formulation, nanocapsules enhanced latanoprost accumulation into the hair follicles, being a nanosystem with high potential for use as a topical formulation for the treatment of androgenic alopecia.

LC-MS bioanalytical method for simultaneous determination of latanoprost and minoxidil in the skin.

The association of minoxidil sulphate and latanoprost is currently emerging as a promising strategy for the treatment of androgenic alopecia, which is the most common type of scalp hair loss. In order to support the development of new pharmaceutical products containing such drugs combination, this study proposes a simple and efficient LC-MS bioanalytical method to simultaneously quantify minoxidil sulphate and latanoprost in different skin layers. Compounds separation was performed by liquid chromatography using a C18 column with gradient elution of a mobile phase composed of 0.1 % formic acid in acetonitrile and water at a flow rate of 0.5 mL min-1. Determinations were executed using mass spectrometry equipped with an ESI interface operating in a positive ionization mode. Quantification was performed using selective ion mode monitoring of m/z 210.1 for minoxidil sulphate and 433.3 for latanoprost. The matrix effect was very pronounced in samples containing some skin layers or electrolyte solution. Accordingly, a calibration curve for each contaminant group was built, leading to correlation coefficient values higher than 0.99. Additionally, lower limits of detection and quantification were obtained, and precision (repeatability and intermediate precision) achieved results with a coefficient of variation less than 15 %. Drug recovery from skin samples was higher than 70 %, fulfilling the recommendations. Also, the bioanalytical method was successfully tested in in vitro skin penetration studies proving its effectiveness in the development of topical formulations containing both drugs.

Microparticles prepared with 50-190kDa chitosan as promising non-toxic carriers for pulmonary delivery of isoniazid.

Chitosan biocompatibility and mucoadhesiveness make it an ideal polymer for antituberculotic drugs microcapsulation for pulmonary delivery. Yet, previous study indicated toxicity problems to J-774.1-cells treated with some medium molecular weight (190-310kDa) chitosan microparticles. As polymer molecular weight is a crucial factor to be considered, this paper describes the preparation and characterization of chitosan (50-190kDa) microparticles containing isoniazid (INH). Cytotoxicity assays were also performed on murine peritoneal (J-774.1) and alveolar (AMJ2-C11) macrophages cell lines, followed by cytokines detection from AMJ2-C11 cells. Spray-drying process produced mucoadhesive microparticles from 3.2µm to 3.9µm, entrapping more than 89% of the drug and preserving their chemical stability. Drug release behavior could be controlled by the use of cross-linked or uncross-linked chitosan, the latter leading to a rapid drug release. Mucoadhesive potential of the microparticles was characterized following in vitro and ex vivo assays. Finally, a significant reduction on toxicity against peritoneal macrophages and no toxic effect on alveolar macrophages with use of such microparticles were observed. In conclusion, 50-190kDa chitosan microparticles may act as promising non-cytotoxic carriers for pulmonary delivery of INH showing marked alveoli macrophage activation.