Compensatory regrowth of the mouse bladder after partial cystectomy.

Cystectomy is the removal of all or part of the urinary bladder. It has been observed that there is significant regrowth of the bladder after partial cystectomy and this has been proposed to be through regeneration of the organ. Regrowth of tissue in mammals has been proposed to involve compensatory mechanisms that share many characteristics of true regeneration, like the growth of specialized structures such as blood vessels or nerves. However, the overall structure of the normal organ is not achieved. Here we tested if bladder growth after subtotal cystectomy (STC, removal of 50% of the bladder) was compensatory or regenerative. To do this we subjected adult female mouse bladders to STC and assessed regrowth using several established cellular parameters including histological, gene expression, cytokine accumulation and cell proliferation studies. Bladder function was analyzed using cystometry and the voiding stain on paper (VSOP) technique. We found that STC bladders were able to increase their ability to hold urine with the majority of volume restoration occurring within the first two weeks. Regenerating bladders had thinner walls with less mean muscle thickness, and they showed increased collagen deposition at the incision as well as throughout the bladder wall suggesting that fibrosis was occurring. Cell populations differed in their response to injury with urothelial regeneration complete by day 7, but stromal and detrusor muscle still incomplete after 8wks. Cells incorporated EdU when administered at the time of surgery and tracing of EdU positive cells over time indicated that many newborn cells originate at the incision and move mediolaterally. Basal urothelial cells and bladder mesenchymal stem cells but not smooth muscle cells significantly incorporated EdU after STC. Since anti-inflammatory cytokines play a role in regeneration, we analyzed expressed cytokines and found that no anti-inflammatory cytokines were present in the bladder 1wk after STC. Our findings suggest that bladder regrowth after cystectomy is compensatory and functions to increase the volume that the bladder can hold. This finding sets the stage for understanding how the bladder responds to cystectomy and how this can be improved in patients after suffering bladder injury.

Tristeza Parasitária em bovinos do semiárido pernambucano / Cattle Tick Fever in semi-arid of Pernambuco

This study aimed to determine the seroprevalence of Babesiosis and Anaplasmosis in cattle from the municipalities of Ouricuri and Petrolina, state of Pernambuco, Brazil, and to define the risk factors for the occurrence of the diseases. Blood samples were collected for serologic testing by Indirect Immunofluorescence Assay (IFA). Sanitary epidemiological questionnaires were applied to the producers aiming to identify possible risk factors. Ticks were collected, identified and tested by Polymerase Chain Reaction (PCR) for the diagnosis of infection by Anaplasma marginale, Babesia bigemina and Babesia bovis. The study was conducted with 861 cattle, being 468 in Petrolina and 393 in Ouricuri. The seroprevalence of A. marginale, B. bigemina and B. bovis in Petrolina was of 35.0% (164/468), 35.9% (168/468) and 32.3% (151/468), respectively; and in Ouricuri was 45.5% (179/393), 38.6% (152/393), and 54.9% (216/393), respectively. Co-infection for Anaplasma spp. and Babesia spp. was observed in 31.6% and 32.1% samples of Petrolina and Ouricuri, respectively. The detection of DNA of Babesia spp. by PCR was possible in 5.8% (8/137) ticks; which 62.5% (5/8) was detected later infection with B. bovis; and 23.3% (32/137) with A. marginale. The presence of ticks, the use of acaricide, age, race, and county of residence of the animals were identified as risk factors for TBD by univariate analysis and multivariate. This study allowed the characterization of the municipalities studied as enzootic instability areas for these hemoparasitic, and consequently alert for adoption of adequate control measures and new studies.

Este estudo objetivou determinar a soroprevalência da Babesiose e Anaplasmose em bovinos dos municípios de Ouricuri e Petrolina, estado de Pernambuco, Brasil; e definir os possíveis fatores de risco para a ocorrência dessas doenças. Amostras de sangue foram coletadas para realização de teste sorológico por Imunofluorescência Indireta (RIFI). Questionários epidemiológicos sanitários foram aplicados aos produtores com o objetivo de identificar possíveis fatores de risco. Carrapatos foram coletados, identificados e testados por Reação em Cadeia da Polimerase (PCR) para o diagnóstico da infecção por Anaplasma marginale, Babesia bigemina e Babaesia bovis. O estudo foi conduzido com 861 bovinos, sendo 468 de Petrolina e 393 de Ouricuri. A soroprevalência de A. marginale, B. bigemina e B. bovis em Petrolina foi de 35,0% (164/468), 35,9% (168/468) e 32,3% (151/468), respectivamente; e em Ouricuri foi de 45,5% (179/393), 38,6% (152/393) e 54,9% (216/393), respectivamente. A co-infecção por Anaplasma spp. e Babesia spp. foi observada em 31,6% e 32,1% de amostras de Petrolina e Ouricuri, respectivamente. A detecção de DNA de Babesia spp. por PCR foi possível em 5,8% (8/137) carrapatos, dos quais em 62,5 % (5/8) foi detectada posteriormente infecção por B. bovis, e em 23,3% (32/137) por A. marginale. A presença de carrapatos, o uso de acaricidas, idade, raça, e o município de residência dos animais foram identificados como fatores de risco para TPB pela análise univariável e multivariável. Este estudo permitiu caracterizar os municípios estudados como de instabilidade enzoótica para esses hemoparasitos, e consequentemente, alertar para adoção de medidas adequadas de controle e realização de novos estudos.

Tristeza Parasitária em bovinos do semiárido pernambucano / Cattle Tick Fever in semi-arid of Pernambuco

Este estudo objetivou determinar a soroprevalência da Babesiose e Anaplasmose em bovinos dos municípios de Ouricuri e Petrolina, estado de Pernambuco, Brasil; e definir os possíveis fatores de risco para a ocorrência dessas doenças. Amostras de sangue foram coletadas para realização de teste sorológico por Imunofluorescência Indireta (RIFI). Questionários epidemiológicos sanitários foram aplicados aos produtores com o objetivo de identificar possíveis fatores de risco. Carrapatos foram coletados, identificados e testados por Reação em Cadeia da Polimerase (PCR) para o diagnóstico da infecção por Anaplasma marginale, Babesia bigemina e Babaesia bovis. O estudo foi conduzido com 861 bovinos, sendo 468 de Petrolina e 393 de Ouricuri. A soroprevalência de A. marginale, B. bigemina e B. bovis em Petrolina foi de 35,0% (164/468), 35,9% (168/468) e 32,3% (151/468), respectivamente; e em Ouricuri foi de 45,5% (179/393), 38,6% (152/393) e 54,9% (216/393), respectivamente. A co-infecção por Anaplasma spp. e Babesia spp. foi observada em 31,6% e 32,1% de amostras de Petrolina e Ouricuri, respectivamente. A detecção de DNA de Babesia spp. por PCR foi possível em 5,8% (8/137) carrapatos, dos quais em 62,5 % (5/8) foi detectada posteriormente infecção por B. bovis, e em 23,3% (32/137) por A. marginale. A presença de carrapatos, o uso de acaricidas, idade, raça, e o município de residência dos animais foram identificados como fatores de risco para TPB pela análise univariável e multivariável. Este estudo permitiu caracterizar os municípios estudados como de instabilidade enzoótica para esses hemoparasitos, e consequentemente, alertar para adoção de medidas adequadas de controle e realização de novos estudos.(AU)

This study aimed to determine the seroprevalence of Babesiosis and Anaplasmosis in cattle from the municipalities of Ouricuri and Petrolina, state of Pernambuco, Brazil, and to define the risk factors for the occurrence of the diseases. Blood samples were collected for serologic testing by Indirect Immunofluorescence Assay (IFA). Sanitary epidemiological questionnaires were applied to the producers aiming to identify possible risk factors. Ticks were collected, identified and tested by Polymerase Chain Reaction (PCR) for the diagnosis of infection by Anaplasma marginale, Babesia bigemina and Babesia bovis. The study was conducted with 861 cattle, being 468 in Petrolina and 393 in Ouricuri. The seroprevalence of A. marginale, B. bigemina and B. bovis in Petrolina was of 35.0% (164/468), 35.9% (168/468) and 32.3% (151/468), respectively; and in Ouricuri was 45.5% (179/393), 38.6% (152/393), and 54.9% (216/393), respectively. Co-infection for Anaplasma spp. and Babesia spp. was observed in 31.6% and 32.1% samples of Petrolina and Ouricuri, respectively. The detection of DNA of Babesia spp. by PCR was possible in 5.8% (8/137) ticks; which 62.5% (5/8) was detected later infection with B. bovis; and 23.3% (32/137) with A. marginale. The presence of ticks, the use of acaricide, age, race, and county of residence of the animals were identified as risk factors for TBD by univariate analysis and multivariate. This study allowed the characterization of the municipalities studied as enzootic instability areas for these hemoparasitic, and consequently alert for adoption of adequate control measures and new studies.(AU)

The Murine Bladder Supports a Population of Stromal Sca-1+/CD34+/lin- Mesenchymal Stem Cells.

Bladder fibrosis is an undesired end point of injury of obstruction and often renders the smooth muscle layer noncompliant. In many cases, the long-term effect of bladder fibrosis is renal failure. Despite our understanding of the progression of this disease, little is known about the cellular mechanisms that lead to a remodeled bladder wall. Resident stem (progenitor) cells have been identified in various organs such as the brain, heart and lung. These cells function normally during organ homeostasis, but become dysregulated after organ injury. Here, we aimed to characterize a mesenchymal progenitor cell population as a first step in understanding its role in bladder fibrosis. Using fluorescence activated cell sorting (FACS), we identified a Sca-1+/ CD34+/ lin- (PECAM-: CD45-: Ter119-) population in the adult murine bladder. These cells were localized to the stromal layer of the adult bladder and appeared by postnatal day 1. Cultured Sca-1+/ CD34+/ lin- bladder cells self-renewed, formed colonies and spontaneously differentiated into cells expressing smooth muscle genes. These cells differentiated into other mesenchymal lineages (chondrocytes, adipocytes and osteocytes) upon culture in induction medium. Both acute and partial obstruction of the bladder reduced expression of CD34 and changed localization of Sca-1 to the urothelium. Partial obstruction resulted in upregulation of fibrosis genes within the Sca-1+/CD34+/lin- population. Our data indicate a resident, mesenchymal stem cell population in the bladder that is altered by bladder obstruction. These findings provide new information about the cellular changes in the bladder that may be associated with bladder fibrosis.

Activity of disinfectants and biofilm production of Corynebacterium pseudotuberculosis / Atividade de desinfetantes e produção de biofilme em Corynebacterium pseudotuberculosis

To verify the occurrence of caseous lymphadenitis in sheep and goats on farms of Pernambuco, Brazil, and in animals slaughtered in two Brazilian cities (Petrolina/PE and Juazeiro/BA), and to characterize the susceptibility profile of Corynebacterium pseudotuberculosis to disinfectants and antimicrobials, and its relationship with biofilm production were the objectives of this study. 398 samples were tested for sensitivity to antimicrobial drugs, disinfectants, and biofilm production. Among the 108 samples collected on the properties, 75% were positive for C. pseudotuberculosis. Slaughterhouse samples indicated an occurrence of caseous lymphadenitis in 15.66% and 6.31% for animals slaughtered in Petrolina and Juazeiro respectively. With respect to antimicrobials, the sensitivity obtained was 100% for florfenicol and tetracycline; 99.25% for enrofloxacin, ciprofloxacin and lincomycin; 98.99% for cephalothin; 98.74% for norfloxacin and sulfazotrim; 97.74% for gentamicin; 94.22% for ampicillin; 91.71% for amoxicillin; 91.21% for penicillin G; 89.19% for neomycin and 0% for novobiocin. In analyzes with disinfectants, the efficiency for chlorhexidine was 100%, 97.20% for quaternary ammonium, 87.40% for chlorine and 84.40% for iodine. 75% of the isolates were weak or non-biofilm producers. For the consolidated biofilm, found that iodine decreased biofilm formation in 13 isolates and quaternary ammonia in 11 isolates. The reduction of the biofilm formation was observed for iodine and quaternary ammonium in consolidated biofilm formation in 33% and 28% of the isolates, respectively. The results of this study highlight the importance of establishing measures to prevent and control the disease.

Verificar a ocorrência de linfadenite caseosa de ovinos e caprinos em fazendas de Pernambuco, no Brasil e em animais abatidos em duas cidades brasileiras (Petrolina/PE e Juazeiro/BA), caracterizar o perfil de suscetibilidade de Corynebacgerium pseudotuberculosis a desinfetantes e a antimicrobianos e analisar sua relação com a produção de biofilme foram os objetivos deste estudo. 398 amostras foram testadas para a sensibilidade aos medicamentos antimicrobianos, desinfetantes e produção de biofilme. Entre as 108 amostras coletadas nas propriedades, 75% foram positivas para C. pseudotuberculosis. Amostras do matadouro indicaram a ocorrência de linfadenite caseosa em 15,66% e 6,31% para animais abatidos em Petrolina (PE) e Juazeiro (BA), respectivamente. Com relação aos antimicrobianos, a sensibilidade obtida foi de 100% para florfenicol e tetraciclina, 99,25% para a enrofloxacina, ciprofloxacina e lincomicina; 98,99% para cefalotina; 98,74% para norfloxacina e sulfazotrim; 97,74% para gentamicina; 94,22% para ampicilina, 91,71% para amoxicilina, 91,21% para a penicilina G, 89,19% para a neomicina e 0% para novobiocina. Nas análises com desinfetantes, a eficiência para a clorexidina foi de 100%, 97,20% para amônia quaternária, 87,40% para o cloro e 84,40% para o iodo. 75% dos isolados foram fracos produtores de biofilme. Para o biofilme consolidado, observou-se que o iodo diminuiu a formação de biofilme em 13 isolados e a amônia quaternária em 11 isolados. A redução da formação de biofilme foi observada no biofilme consolidado, para iodo e amônia quaternária, em 33% e 28% das amostras, respectivamente. Os resultados deste estudo destacam a importância de se estabelecer medidas para prevenir e controlar a doença.

Binding to and photo-oxidation of cardiolipin by the phthalocyanine photosensitizer Pc 4.

Cardiolipin is a unique phospholipid of the mitochondrial inner membrane. Its peroxidation correlates with release of cytochrome c and induction of apoptosis. The phthalocyanine photosensitizer Pc 4 binds preferentially to the mitochondria and endoplasmic reticulum. Earlier Förster resonance energy transfer studies showed colocalization of Pc 4 and cardiolipin, which suggests cardiolipin as a target of photodynamic therapy (PDT) with Pc 4. Using liposomes as membrane models, we find that Pc 4 binds to cardiolipin-containing liposomes similarly to those that do not contain cardiolipin. Pc 4 binding is also studied in MCF-7c3 cells and those whose cardiolipin content was reduced by treatment with palmitate. Decreased levels of cardiolipin are quantified by thin-layer chromatography. The similar level of binding of Pc 4 to cells, irrespective of palmitate treatment, supports the lack of specificity of Pc 4 binding. Thus, factors other than cardiolipin are likely responsible for the preferential localization of Pc 4 in mitochondria. Nonetheless, cardiolipin within liposomes is readily oxidized by Pc 4 and light, yielding apparently mono- and dihydroperoxidized cardiolipin. If similar products result from exposure of cells to Pc 4-PDT, they could be part of the early events leading to apoptosis following Pc 4-PDT.