Dedifferentiation of prostate smooth muscle cells in response to bacterial LPS

Prostate smooth muscle cells (SMCs) are strongly involved in the development and progression of benign prostatic hyperplasia and prostate cancer. However, their participation in prostatitis has not been completely elucidated. Thus, we aimed to characterize the response of normal SMC to bacterial lipopolysaccharide (LPS). Methods Primary prostate SMCs from normal rats were stimulated with LPS (0.1, 1, or 10Âμg/ml) for 24 or 48hr. The phenotype was evaluated by electron microscopy, immunofluorescence, and Western blot of SMCα-actin (ACTA2), calponin, vimentin, and tenascin-C, while the innate immune response was assessed by immunodetection of TLR4, CD14, and nuclear NF-κB. The secretion of TNFα and IL6 was determined using ELISA. Results Bacterial LPS induces SMCs to develop a secretory phenotype including dilated rough endoplasmic reticulum cisternae with well-developed Golgi complexes. Furthermore, SMCs displayed a decrease in ACTA2 and calponin, and an increase in vimentin levels after LPS challenge. The co-expression of ACTA2 and vimentin, together with the induction of tenascin-C expression indicate that a myofibroblastic-like phenotype was induced by the endotoxin. Moreover, LPS elicited a TLR4 increase, with a peak in NF-κB activation occurring after 10min of treatment. Finally, LPS stimulated the secretion of IL6 and TNFα. ConclusionS Prostate SMCs are capable of responding to LPS in vitro by dedifferentiating from a contractile to a miofibroblastic-like phenotype and secreting cytokines, with the TLR4 signaling pathway being involved in this response. In this way, prostate SMCs may contribute to the pathophysiology of inflammatory diseases by modifying the epithelial-stromal interactions.Fil: Leimgruber, Carolina. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Ciencias de la Salud. Universidad Nacional de Córdoba. Instituto de Investigaciones en Ciencias de la Salud; ArgentinaFil: Quintar, Amado A.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Ciencias de la Salud. Universidad Nacional de Córdoba. Instituto de Investigaciones en Ciencias de la Salud; Argentina. Universidad Nacional de Córdoba; ArgentinaFil: Sosa, Liliana del Valle. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Ciencias de la Salud. Universidad Nacional de Córdoba. Instituto de Investigaciones en Ciencias de la Salud; ArgentinaFil: García, Luciana Noemí. Universidad Nacional de Córdoba; Argentina. Centro de Investigación de la Fundación Repro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Figueredo, Carlos Mauricio. Universidad Nacional de Córdoba; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Maldonado, Cristina Alicia. Universidad Nacional de Córdoba. Facultad de Medicina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Ciencias de la Salud. Universidad Nacional de Córdoba. Instituto de Investigaciones en Ciencias de la Salud; Argentin

Ectoparasite infestation on rural dogs in the municipality of São Vicente Férrer, Pernambuco, Northeastern Brazil

This article presents the results of a study on ectoparasites infesting rural dogs in Pernambuco, northeastern Brazil. Forty-one dogs from a rural community of Pernambuco were examined and 70.7% were infested by ectoparasites. The prevalence rates of infestation by ticks, fleas, and lice were 58.5, 43.9 and 22%, respectively. Of 24 dogs parasitized by ticks, 15 were exclusively infested by Rhipicephalus sanguineus (Latreille) (32 females, 66 males, 9 nymphs; prevalence, 48.8%; mean intensity, 5.4), four by Amblyomma ovale Koch (33 females, 19 males; prevalence, 22%; mean intensity, 5.8), and five were coinfested by both species. Ctenocephalides felis felis (Bouché) (25 females, 8 males; prevalence, 43.9%; mean intensity, 1.9) and Heterodoxus spiniger (Enderlein) (16 females, 11 males, 10 nymphs; prevalence, 22%; mean intensity, 4.1) were the only species of flea and louse identified. Tick infestation was more frequent than flea or lice infestations (p < 0.05). No statistical difference was found by sex and age group of the dogs studied and prevalence of infestation. Overall, 48.8% (20/41) of the dogs were infested by more than one ectoparasite species

Antifungal susceptibility of Malassezia pachydermatis

Antifungal resistance has been associated with biofilm formation in many microorganisms, but not yet in Malassezia pachydermatis. This saprophytic yeast can cause otitis and dermatitis in dogs and has emerged as an important human pathogen, responsible for systemic infections in neonates in intensive care units. This study aims to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains, in both their planktonic and sessile forms, to fluconazole, miconazole, ketoconazole, itraconazole, posaconazole, terbinafine and voriconazole using the XTT assay and Clinical and Laboratory Standards Institute (CLSI) microdilution method. The minimum inhibitory concentration (MIC) values recorded for each drug were significantly higher for sessile cells relative to planktonic cells to the extent that ≥ 90% of M. pachydermatis strains in their sessile form were classified as resistant to all antifungal agents tested. Data suggest that M. pachydermatis biofilm formation is associated with antifungal resistance, paving the way towards investigating drug resistance mechanisms in Malassezia spp

Transovarial passage of Leishmania infantum kDNA in artificially infected Rhipicephalus sanguineus

Submitted by Kamylla Nascimento ([email protected]) on 2018-08-29T13:22:58Z No. of bitstreams: 1 Transovarial passage of Leishmania infantum kDNA in artificially infected.pdf: 114960 bytes, checksum: a15764cf775561b6a73eb082fb9092f1 (MD5)Approved for entry into archive by Kamylla Nascimento ([email protected]) on 2018-08-29T13:30:41Z (GMT) No. of bitstreams: 1 Transovarial passage of Leishmania infantum kDNA in artificially infected.pdf: 114960 bytes, checksum: a15764cf775561b6a73eb082fb9092f1 (MD5)Made available in DSpace on 2018-08-29T13:30:41Z (GMT). No. of bitstreams: 1 Transovarial passage of Leishmania infantum kDNA in artificially infected.pdf: 114960 bytes, checksum: a15764cf775561b6a73eb082fb9092f1 (MD5) Previous issue date: 2010Università degli Studi di Bari. Dipartimento di Sanità Pubblica e Zootecnia. Valenzano, BA, Italy.Universidade de São Paulo. Departamento de Medicina Veterinária Preventiva e Saúde Animal. São Paulo, SP, Brazil.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil.Università degli Studi di Bari. Dipartimento di Sanità Pubblica e Zootecnia. Valenzano, BA, Italy.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Imunologia. Recife, PE, Brasil.Phlebotomine sand flies are the only proven biological vectors of Leishmania parasites. However, Rhipicephalus sanguineus ticks have long been suspected to transmit Leishmania infantum in studies carried out in laboratory and natural conditions. In the present study, 5 microl of L. infantum promastigotes (1x10(6) cells per ml) was injected into the hemocel through the coxa I of four engorged females (F1, F2, F3 and F4). Control ticks (F5 and F6) were injected with sterile phosphate-buffered saline (PBS) using the same procedure. Then, these females, their eggs, and the originated larvae were tested by real time polymerase chain reaction (real-time PCR) for the presence of L. infantum kinetoplast DNA (kDNA). Females and eggs were tested after the end of the oviposition period (about 5 weeks post-inoculation) whereas larvae were tested about 4 months after the inoculation of females. All artificially infected females were positive for L. infantum kDNA. In addition, two pools of eggs (one from F2 and other from F4) and four pools of larvae (one from each F1 and F4 and two from F2) were positive for L. infantum kDNA. These results showed, for the first time, the transovarial passage of L. infantum kDNA in R. sanguineus ticks, thus suggesting that the transovarial transmission of L. infantum protozoa in ticks is worth to be investigated

Fusarium spp. in Loggerhead Sea Turtles (Caretta caretta): From Colonization to Infection

With the aim of evaluating the presence of Fusarium spp. in sea turtles with and without lesions and assessing the risk factors favoring colonization and/or infection, 74 loggerhead sea turtles (Caretta caretta) admitted to rescue and rehabilitation clinics in Italy were analyzed. The study compared 31 individuals with no apparent macroscopic lesions and 43 individuals with macroscopic lesions. Shell and skin samples were analyzed using Calcofluor white with 10% potassium hydroxide, standard histopathological examination, and fungal cultures. Fusarium spp. were isolated more frequently from animals with superficial lesions (39%) than from those with no macroscopic lesions (16%). Isolates from animals with superficial lesions were Fusarium solani species complex (FSSC) lineages haplotypes 9, 12, and 27 (unnamed lineages), FSSC-2 (Fusarium keratoplasticum), Fusarium oxysporum (27%), and Fusarium brachygibbosum (3%). In contrast, only F. solani haplotypes 9 and 12 were isolated from animals with no macroscopic lesions. The presence of lesions was identified as a risk factor for the occurrence of Fusarium spp. Of the 74 animals, only 7 (9.5%) scored positive on microscopic examination with Calcofluor, and histological examination of those 7 animals revealed necrosis, inflammatory cells, and fungal hyphae in the carapace and skin. The results of this study suggest that fusariosis should be included in the differential diagnosis of shell and skin lesions in sea turtles. Direct examination using Calcofluor and potassium hydroxide was not useful to diagnose the infection. Histopathological examination and fungal culture should be performed to ensure correct treatment and infection control

Fusarium spp. in Loggerhead Sea Turtles (Caretta caretta): From Colonization to Infection

With the aim of evaluating the presence of Fusarium spp. in sea turtles with and without lesions and assessing the risk factors favoring colonization and/or infection, 74 loggerhead sea turtles (Caretta caretta) admitted to rescue and rehabilitation clinics in Italy were analyzed. The study compared 31 individuals with no apparent macroscopic lesions and 43 individuals with macroscopic lesions. Shell and skin samples were analyzed using Calcofluor white with 10% potassium hydroxide, standard histopathological examination, and fungal cultures. Fusarium spp. were isolated more frequently from animals with superficial lesions (39%) than from those with no macroscopic lesions (16%). Isolates from animals with superficial lesions were Fusarium solani species complex (FSSC) lineages haplotypes 9, 12, and 27 (unnamed lineages), FSSC-2 (Fusarium keratoplasticum), Fusarium oxysporum (27%), and Fusarium brachygibbosum (3%). In contrast, only F. solani haplotypes 9 and 12 were isolated from animals with no macroscopic lesions. The presence of lesions was identified as a risk factor for the occurrence of Fusarium spp. Of the 74 animals, only 7 (9.5%) scored positive on microscopic examination with Calcofluor, and histological examination of those 7 animals revealed necrosis, inflammatory cells, and fungal hyphae in the carapace and skin. The results of this study suggest that fusariosis should be included in the differential diagnosis of shell and skin lesions in sea turtles. Direct examination using Calcofluor and potassium hydroxide was not useful to diagnose the infection. Histopathological examination and fungal culture should be performed to ensure correct treatment and infection control.</p

In vitro

Dermatitis caused by Malassezia spp., one of most common skin disease in dogs, requires prolonged therapy and/or high doses of antifungal agents. In the present study, the antifungal susceptibility of M. pachydermatis to ketoconazole (KTZ), fluconazole (FLZ), itraconazole (ITZ), posaconazole (POS) and voriconazole (VOR) was evaluated in vitro using both CLSI reference broth microdilution (CLSI BMD) and E-test. A total of 62 M. pachydermatis strains from dogs with and without skin lesions were tested. M. pachydermatis strains were susceptible to ITZ, KTZ and POS using both test methods, with the highest MIC found in tests of FLZ. Essential agreement between the two methods ranged from 87.1% (VOR) to 91.9% (ITZ), and categorical agreement from 74.2% (FLZ) to 96.8% (ITZ). Minor error discrepancies were observed between the two methods, with major discrepancies observed for KTZ. A higher MIC(50) value for FLZ was noted with M. pachydermatis genotype B. The MICs(50) of M. pachydermatis genotype B for KTZ, VOR and POS were higher in isolates from dogs with skin lesions than those in isolates from animals without skin lesions. The results suggest a link between genotypes of M. pachydermatis and in vitro drug susceptibility. The categorical agreement for both E-test and CLSI BMD methods found in this investigation confirms the E-test as a reliable diagnostic method for routine use in clinical mycology laboratories