Antifungal activity of 2-chloro-N-phenylacetamide: a new molecule with fungicidal and antibiofilm activity against fluconazole-resistant Candida spp

Abstract In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

Resumo No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Antifungal activity of 2-chloro-N-phenylacetamide: a new molecule with fungicidal and antibiofilm activity against fluconazole-resistant Candida spp / Atividade antifúngica de 2-cloro-N-fenilacetamida: uma nova molécula com atividade fungicida e anti-biofilme contra Candida spp. resistentes a fluconazol

In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

No atual contexto de patógenos fúngicos resistentes emergentes tais como Candida albicans e Candida parapsilosis, a descoberta de novos agentes antifúngicos é uma questão urgente. Esta pesquisa teve como objetivo avaliar o potencial antifúngico da 2-cloro-N-fenilacetamida contra cepas clínicas de C. albicans e C. parapsilosis resistentes a fluconazol. A atividade antifúngica da substância foi avaliada in vitro através da determinação da concentração inibitória mínima (CIM), concentração fungicida mínima (CFM), ruptura e inibição da formação de biofilme, ensaios de sorbitol e ergosterol, e associação entre esta molécula e antifúngicos comuns, anfotericina B e fluconazol. O produto teste inibiu todas as cepas de C. albicans e C. parapsilosis, com uma CIM variando de 128 a 256 µg.mL-1, e uma CFM de 512-1,024 µg.mL-1. Também inibiu até 92% da formação de biofilme e causou a ruptura de até 87% de biofilme pré-formado. A 2-cloro-N-fenilacetamida não promoveu atividade antifúngica pela ligação ao ergosterol da membrana celular fúngica, tampouco danificou a parede celular. Antagonismo foi observado ao combinar esta substância com anfotericina B e fluconazol. A substância exibiu atividade antifúngica significativa ao inibir tanto as células planctônicas quanto o biofilme das cepas resistentes ao fluconazol. Sua combinação com outros antifúngicos deve ser evitada e seu mecanismo de ação deve ser estabelecido.

Evaluation of the antimicrobial effect of the Origanum vulgare L essential oil on strains of Klebsiella pneumoniae.

Bacteria may be the initial cause of certain pathologies as well as a secondary agent responsible for the development of complications such as pressure ulcer infections. Pressure ulcers are a persistent health problem, especially in immunocompromised patients, and associated with infection by opportunistic microorganisms with antimicrobial resistance, such as Klebsiella pneumoniae, highlight the need for the development of new antimicrobial approaches. Thus, the aim of this study was to evaluate the antibacterial and anti-adherent activity of Origanum vulgare L. (oregano) essential oil against Klebsiella pneumoniae strains, as well as the effect of its association with synthetic antimicrobials. To this end, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) analyses were performed on microdilution plates. The assay of the Minimum Inhibitory Adherence Concentration (MIAC), with test tubes. As well as, the association study through the infusion disc method containing ampicillin (AMP), gentamicin (GEN), ciprofloxacin (CIP) and ceftriaxone (CEF). Therefore, it was possible to obtain that the essential oil of oregano presents antimicrobial and bactericidal activity, with MIC ranging between 128µg/mL and 256 µg/mL and MBC between 256 µg/mL and 512 µg/mL, on the tested K. pneumoniae strains. When used in association with ampicillin and gentamicin, oregano essential oil showed synergistic effect for some strains. Therefore, it is observed that the tested essential oil can act as a promising antibacterial in the treatment of diseases caused by K. pneumoniae.

Hereditary angioedema: 24 years of experience in a Portuguese Reference Center.

Summary: Hereditary angioedema (HAE) poses a high burden of disease, being its epidemiological and clinical data heterogeneous among countries, with no recent published studies concerning Portuguese patients. Therefore, we aimed to raise awareness of HAE and to contribute to clinical knowledge. An observational, descriptive, retrospective, and cross-sectional study was performed, that included a cohort of 126 patients followed in a single Portuguese Center. We observed a high prevalence of HAE-C1-INH type II (45.2% of patients). Most HAE patients (67.4%) presented the initial manifestations of the disease before adulthood, at a mean age of 12.6 ± 8.4 years. However, we found a long delay in HAE diagnosis, especially in those without family history (mean 20.7 ± 17.3 years). Stress was the most common trigger, followed by trauma and infection. Symptoms involving different systems were increasingly reported with increased disease duration. Cutaneous symptoms (95.0%) were more frequent, followed by gastrointestinal (80.7%), and respiratory symptoms (50.4%). HAE symptoms led to abdominal surgery in 22 (17.5%) patients and induced laryngeal edema requiring intubation/tracheostomy in 8 (6.3%) patients. Most patients were under long-term prophylaxis, mainly with attenuated androgens (62.7% of patients).The correct distinction between HAE and other common causes of angioedema is critical, allowing reduction of diagnostic delay, improvement of adequate management, and ultimately improving outcomes and quality of life of HAE patients.

Effects of canola oil and antioxidants on performance, serum parameters, carcass traits, and rumen fermentation patterns of Nellore cattle.

Several nutritional strategies have been used in beef cattle production in order to increase animal performance and profitability. However, in the past two decades, the increase of consumer preference for functional foods has driven the investigation for improving food via adding functional substances to animal diets. We evaluated the effect of canola oil supplementation associated with vitamin E and selenium on performance, rumen metabolism, carcass traits, meat tenderness, and serum, liver, and meat status of antioxidants in finishing Nellore males. Animals were fed for 106 days in a feedlot and were randomly distributed in a 2 × 2 factorial arrangement: two levels of oil in the diet (no inclusion and 3% canola oil, defined as diet without oil inclusion (NO) and effect of oil (OIL), respectively) and two levels of antioxidants in the diet (no inclusion and 2.5 mg of Se/kg of DM + 500 UI of vitamin E/kg of DM, defined as diet without antioxidant inclusion (NA) and effect of the antioxidants (ANT), respectively). DM intake (kg/day) was evaluated daily; performance and serum were analysed at the beginning of the feedlot and every 28 days. Animals were slaughtered and hot carcass weight (kg) was recorded; ruminal fluid and liver samples were collected. At 24 h postmortem, carcass pH was recorded and the Longissimus thoracis was sampled. There was no significant effect of the OIL*ANT interaction (P > 0.05) for any trait evaluated. Bulls fed OIL presented greater final BW (P < 0.01), average daily gain (kg/day; P < 0.01), feed efficiency (P < 0.01), rump fat thickness (P8RF; P < 0.05), and greater tenderness; the ANT diet increased P8RF (P < 0.05). The levels of selenium and vitamin E in serum, liver, and meat were increased (P < 0.01) with the inclusion of ANT. ANT did not change triiodothyronine (T3, ng/mL) and thyroxine (T4, µg/gL) serum concentrations but decreased serum glucose levels. The treatments did not affect (P > 0.05) ruminal parameters or the protozoa population. Our results showed that the inclusion of 3% canola oil in the diet DM increased performance, feed efficiency, carcass fat deposition, and tenderness, with no effect on rumen fermentation and protozoa population of Nellore cattle in a feedlot system. The inclusion of ANT in the cattle diet did not affect performance or rumen parameters. However, the levels of ANT were increased in the serum, liver, and meat, enriching the final product with these compounds.

Nutritional value, total dry matter losses, effluent production and pollutant potential in Brachiaria brizantha cv. Paiaguás grass / [Valor nutricional, perdas totais de matéria seca, produção e potencial poluidor de efluentes em silagem de Brachiaria brizantha cv. Paiaguás]

The objective of the present study was to evaluate losses, production and polluting potential of the effluent, nutritional value and aerobic stability of silages of Brachiaria brizantha cv. Paiaguás grass, in different particle sizes and compaction density in silage. Three theoretical particle sizes (TTP 5; 8 and 12mm) and three compaction densities (DC 550; 600 and 650kg/m3) were evaluated, distributed in a factorial design (3 x 3), with four repetitions. The highest volume of effluent was found in silages with higher compaction densities (600 and 650kg/m3) and lower TTP (5 and 8mm). The highest chemical oxygen demand and biochemical oxygen demand were registered in the treatment with TTP of 5mm and higher DC (600 and 650kg/m3). Greater in vitro digestibility of DM was verified in the silage chopped at 5 and 8mm. There was no break in aerobic stability for 216 hours. Silage with a low compaction density 550kg/m3 and processing with a theoretical particle size of 12mm reduces effluent losses. In general, the nutritional value of Paiaguás grass was not influenced by the treatments. Different particle sizes and compaction density did not change the aerobic stability of silages.(AU)

Objetivou-se avaliar perdas, produção e potencial poluidor do efluente, valor nutricional e estabilidade aeróbia de silagens do capim Brachiaria brizantha cv. Paiaguás, em diferentes tamanhos de partícula e densidade de compactação na ensilagem. Foram avaliados três tamanhos teóricos de partícula (TTP 5; 8 e 12mm) e três densidades de compactação (DC 550; 600 e 650kg/m3), distribuídos em arranjo fatorial (3 x 3), com quatro repetições. O maior volume de efluente foi verificado nas silagens com maiores densidades de compactação (600 e 650kg/m3) e menores TTP (5 e 8mm). As maiores demanda química de oxigênio e demanda bioquímica de oxigênio foram registradas no tratamento com TTP de 5mm e nas maiores DC (600 e 650kg/m3). Maior digestibilidade in vitro da MS (média de 57,2%) foi verificada na silagem picada a 5 e 8mm. Não houve quebra da estabilidade aeróbia durante 216 horas. A ensilagem com baixa densidade de compactação (550kg/m3) e o processamento com tamanho teórico de partículas 12mm reduzem as perdas por efluente. O valor nutricional da silagem de capim-paiaguás, em geral, não foi influenciado pelos tratamentos. Diferentes tamanhos de partícula e densidade de compactação não alteraram a estabilidade aeróbia das silagens.(AU)

Fine needle aspiration cytology in feline skeletal muscle as a diagnostic tool for extramedullary plasmacytoma / Citologia aspirativa por agulha fina no músculo esquelético felino como ferramenta diagnóstica para o plasmocitoma extramedular

Extramedullary noncutaneous plasmacytoma (ENP) is a myeloproliferative disorder of plasma cells that rarely affects cats. This paper describes an ENP case revealed by fine needle aspiration cytology (FNAC) of the mass in the skeletal muscle of an 8-month-old, male, mixed breed cat, which had a nodule in the left hind limb. The rapid immunoassay test confirmed the presence of feline leukemia virus (FeLV). The animal necropsy macroscopically showed the nodule came from the semimembranosus muscle. Histopathological examination ratified the cytological findings. Thus, this paper alerts to the existence of plasmacytoma located in the skeletal muscle of feline species. FNAC is a quick and efficient method for diagnosis of ENP.(AU)

O plasmocitoma extramedular (PEM) não cutâneo é um distúrbio mieloproliferativo de plasmócitos que raramente acomete felinos. Este trabalho descreve um caso de PEM no músculo esquelético de um gato, macho, sem raça definida, de oito meses de idade, que apresentava um aumento de volume no membro pélvico esquerdo. A citologia aspirativa por agulha fina (CAAF) da massa revelou tratar-se de PEM. O teste imunoenzimático rápido confirmou a presença do vírus da leucemia felina (FeLV). Na necropsia do animal, macroscopicamente, percebeu-se que o nódulo era originário do músculo semimembranoso. O exame histopatológico ratificou os achados citológicos. Desta forma, alerta-se para a existência de plasmocitoma com localização em músculo esquelético na espécie felina, sendo a CAAF um método alternativo rápido e eficaz para o seu diagnóstico.(AU)

Effects of immunocastration and ß-adrenergic agonists on the performance and carcass traits of feedlot finished Nellore cattle.

ß-Adrenergic agonists (ß-AA) are non-hormonal growth promoters which promote muscle hypertrophy in supplemented animals. The effects of two ß-AA in combination with the immunocastration technique on the performance and carcass traits were evaluated using 96 feedlot Nellore males in a randomized complete block design with two sex conditions (immunocastrated (IC) v. non-castrated (NC)) and three treatments: CON (no ß-agonists added), RH (300 mg of ractopamine hydrochloride/day, for 33 days) or ZH (80 mg of zilpaterol·hydrochloride animal/day for 30 days, removed 3 days for required withdrawal period). The trial was carried for 100 days where in the first 70 days animals did not receive ß-AA (phase 1) and during the last 30 days they were treated with ß-AA (phase 2). The performance and ultrasound measurements of longissimus muscle area (LMA), backfat thickness (BFT) and rump fat thickness (RFT) were evaluated in both phases. No sex condition v. treatment interactions were observed for any trait. The NC animals had higher average daily gain (ADG) and final BW than the IC animals, but they did not differ in dry matter intake (DMI) and feed efficiency (gain to feed). The NC animals showed greater LMA (P=0.0001) and hot carcass weight (P=0.0006), and smaller BFT (P=0.0007), RFT (P=0.0039) and percentage of kidney, pelvic and heart fat (P<0.0001) when compared with IC animals. The animals fed ZH showed greater ADG (P=0.0002), G : F (P<0.0001) and dressing per cent (P=0.0136) than those fed RH and CON diets. No differences in BW and DMI were observed. A interaction between treatment and time on feed was observed for LMA and BFT, in which the animals fed ZH diet showed greater LMA (P<0.01) and lower BFT (P<0.01) at 100 days than the animals fed RH and CON diets, whereas RH and CON diets did not differ. Immunocastration decreases muscle development and increases carcass finishing. In contrast, ß-AA increases muscle and decreases fat deposition. The ZH has a higher action on the muscle metabolism than animals fed RH diet. However, RH diet achieves a better balance because it has an intermediary performance between non-supplemented and ZH animals and does not decrease the carcass fat.

Toxicological Evaluation of Essential Oil From the Leaves of Croton argyrophyllus (Euphorbiaceae) on Aedes aegypti (Diptera: Culicidae) and Mus musculus (Rodentia: Muridae).

Plant-derived essential oils can be used as insecticides for vector control. However, to establish their safety, it is necessary to perform toxicological studies. Herein, we evaluated the chemical composition and insecticidal activity of the essential oil from the leaves of Croton argyrophyllus on the third- and fourth-instar larvae and adult Aedes aegypti (L., 1762). We also evaluated the acute toxicity of the essential oil in adult female Mus musculus. The lethal concentration 50 (LC50) and 90 (LC90) of C. argyrophyllus essential oil on larvae of Ae. aegypti were 0.31 and 0.70 mg ml-1, respectively, and 5.92 and 8.94 mg ml-1, respectively, on Ae. aegypti adults. The major components of the essential oil were spathulenol (22.80%), (E)-caryophyllene (15.41%), α-pinene (14.07%), and bicyclogermacrene (10.43%). It also displayed acute toxicity in adults of Mus musculus; the intraperitoneal and oral lethal dose 50 (LD50) were 2,000 mg kg-1 and 2,500 mg kg-1, respectively. The results showed that the essential oil from C. argyrophyllus leaves has insecticidal activity on Ae. aegypti larvae and adults at an average lethal concentration below the median lethal dose needed to cause acute toxicity in the common mouse.

Snake Venom Peptides and Low Mass Proteins: Molecular Tools and Therapeutic Agents.

Snake venoms are natural sources of biologically active molecules that are able to act selectively and specifically on different cellular targets, modulating physiological functions. Thus, these mixtures, composed mainly of proteins and peptides, provide ample and challenging opportunities and a diversified molecular architecture to design and develop tools and agents of scientific and therapeutic interest. Among these components, peptides and small proteins play diverse roles in numerous physiological processes, exerting a wide range of pharmacological activities, such as antimicrobial, antihypertensive, analgesic, antitumor, analgesic, among others. The pharmaceutical and cosmetic industries have recognized the huge potential of these privileged frameworks and believe them to be a promising alternative to contemporary drugs. A number of natural or synthetic peptides from snake venoms have already found preclinical or clinical applications for the treatment of pain, hypertension, cardiovascular diseases and aging skin. A well-known example is captopril, whose natural peptide precursor was isolated from Bothrops jararaca snake venom, which is a peptide-based drug that inhibits the angiotensin-converting enzyme, producing an anti-hypertensive effect. The present review looks at the main peptides (natriuretic peptides, bradykinin-potentiating peptides and sarafotoxins) and low mass proteins (crotamine, disintegrins and three-Finger toxins) from snake venoms, as well as synthetic peptides inspired by them, describing their biochemical, structural and physiological features, as well as their applications as research tools and therapeutic agents.

Artery Stenosis of the Renal Graft: Experience of a Center of Northeastern Brazil.

BACKGROUND: Transplant renal artery stenosis (TRAS), the most common vascular complication after transplant (Tx), leads to resistant hypertension, impaired renal function, and even loss of the graft. The purpose of the study was to investigate the prevalence and factors associated with TRAS in northeastern Brazil. METHODS: The study was conducted as a retrospective case-control study in a population of Tx recipients in a renal Tx center in northeastern Brazil. Demographic and clinical characteristics of the recipients and donors, data related to the surgery, laboratory data, and number of anti-hypertensive drugs were assessed. Statistical analysis was performed with the use of SPSS 17.0. RESULTS: A total of 494 of 529 recipients were assessed, of which 24 had TRAS. The prevalence of TRAS was 4.8%. Twelve patients (50%) were men with a mean age of 46.7 ± 13.5 years. The mean time of diagnosis was 89.9 days after Tx. The risk factors associated with TRAS were number of anti-hypertensive drugs ≥2 (odds ratio, 17.0; confidence interval, 4.1 to 70.4; P = .001) and grafting with 2 or more arteries (odds ratio, 8.9; confidence interval, 1.4 to 56.6; P = .021). There was a significant reduction in mean systolic blood pressure (147.1 ± 23.7 to 127.8 ± 15.2 mm Hg, P = .001) and diastolic blood pressure (86.6 ± 13.0 to 77.6 ± 9.4 mm Hg, P = .001) after TRAS repair and in serum creatinine (2.8 ± 2.4 to 1.9 ± 1.8 mg/dL, P = .04). CONCLUSIONS: Grafts with 2 or more arteries are associated with TRAS, as well as patients who use a higher number of anti-hypertensive drugs. TRAS repair was associated with improved blood pressure control and renal function.

Severe bronchiectasis in a patient with common variable immunodeficiency.

BACKGROUND: Bronchiectasis are common in Common Variable Immunodeficiency. These patients are prone to infection, leading to progressive lung destruction and accelerated FEV1 decline. CLINICAL CASE: 40 year-old man, with recurrent respiratory infections, autoimmunity and diarrhea since age 7. At 17 CVID was diagnosed and IVIgG was started. During the following years, respiratory symptoms progressively worsened and bronchiectasis was found on thoracic computed tomography. Bronchoscopy revealed Pseudomonas aeruginosa in bronchoalveolar lavage and bronchial secretions cultures. Eradication therapy led to clinical improvement. DISCUSSION: This case report stresses the importance of regular microbiological screening and appropriate antibiotherapy. Early/aggressive treatment may significantly impact on patients' evolution.

Monocyte activation is a feature of common variable immunodeficiency irrespective of plasma lipopolysaccharide levels.

Common variable immunodeficiency disorders (CVID), the most frequent cause of symptomatic primary immunodeficiency, are defined by impaired antibody production. Notwithstanding, T cell activation and granulomatous manifestations represent the main causes of CVID morbidity even in patients receiving immunoglobulin (Ig) G replacement therapy. Additionally, gut pathology is a frequent feature of CVID. In this study, we investigated monocyte imbalances and their possible relationship with increased microbial translocation in CVID patients. Monocyte subsets were defined according to CD14 and CD16 expression levels and evaluated in terms of human leucocyte antigen D-related (HLA-DR), CD86 and programmed death-1 molecule ligand 1 (PD-L1) expression by flow cytometry, in parallel with the quantification of plasma lipopolysaccharide (LPS) and serum levels of soluble CD14 (sCD14), LPS-binding protein (LBP) and anti-LPS antibodies. CVID patients (n=31) featured significantly increased levels of serum sCD14 and an expansion of CD14(bright) CD16(+) monocytes in direct correlation with T cell and B cell activation, the latter illustrated by the frequency of the CD21(low) CD38(low) subset. Such alterations were not observed in patients lacking B cells due to congenital agammaglobulinaemia (n=4). Moreover, we found no significant increase in circulating LPS or LBP levels in CVID patients, together with a relative preservation of serum anti-LPS antibodies, in agreement with their presence in commercial IgG preparations. In conclusion, CVID was associated with monocyte imbalances that correlated directly with T cell activation markers and with B cell imbalances, without an association with plasma LPS levels. The heightened monocyte activated state observed in CVID may represent an important target for complementary therapeutic strategies.

Polyomavirus-infected decoy cells in cytocentrifuged urine cytology specimens from renal transplant recipients.

OBJECTIVE: To evaluate the prevalence of BK virus (BKV) infection in renal transplant recipients based on the detection of urinary decoy cells. METHODS: Fifty transplantation patients (56% males) aged 39.9 ± 11 years were screened for urinary decoy cells. The majority (86%) had received grafts from living donors. The inclusion criteria were graft dysfunction, hematuria, and/or leukocyturia. Creatinine and urea serum levels were monitored for 6 months. RESULTS: Decoy cells were found in the urine of 12 (24%) patients 1-2 years after transplantation. The immunosuppression regimens most frequently adopted by BKV-positive patients were cyclosporine + azathiprine (50%) and FK + mycophenolate mofetyl + prednisone (25%). A renal biopsy revealed normal structures in 7 patients (58.4%), BKV nephropathy with lymphomononuclear infiltrate and fibrosis in 4 patients (33.3%), and acute cellular rejection with lymphomononuclear infiltrate without fibrosis in 1 patient (8.3%). In the latter patient, BKV-associated nephropathy was confirmed by immunohistochemistry. During the first month, creatinine and urea serum levels were higher among patients with urinary decoy cells. Creatinine levels decreased between the first and the sixth months. CONCLUSION: The finding of BKV-associated nephropathy in 33.3% of the patients with urinary decoy cells stresses the importance of screening for BKV with urinary cytology.

Carcass quality of feedlot finished steers fed yeast, monensin, and the association of both additives / Qualidade de carcaça de novilhos terminados com dietas contendo levedura, monensina e associação de ambos aditivos

To evaluate the effects of the supplementation of feed additives on carcass quality in beef cattle, 72 Nellore steers (339.5kg, 20-month old) were feedlot finished and fed for 91 days one of the following diets: 1) control with no additives; or added of 2) live yeast culture; 3) monensin; or 4) the association of both additives. After slaughter, renal, pelvic, and inguinal fat and hot carcass weights were recorded and carcass was split into muscle, bone, and trimmable fat. Carcass Longissimus muscle area and subcutaneous fat thickness at the 12th rib were measured and steaks of Longisimus muscle were taken to determine meat color, shear force, drip, and cooking losses. Yeast increased carcass dressing percentage but there were no effects on hot carcass weight, Longissimus area, subcutaneous fat thickness, percentage and weight of retail cut yield and trimmings. Feed additives had no effect on carcass pH, meat color, fat content, shear force, and drip losses. Supplementation of yeast, monensin or the association of both additives had no important effects on carcass traits and on meat quality of feedlot finished steers.

Avaliaram-se os efeitos da suplementação de aditivos alimentares sobre a qualidade de carcaça em bovinos de corte. Usaram-se 72 novilhos Nelore com média de peso de 339,5kg e 20 meses de idade, terminados em confinamento e alimentados por 91 dias com uma das quatro dietas: 1) dieta controle sem aditivos, ou com a adição de 2) leveduras vivas, 3) monensina ou 4) associação entre ambos aditivos. Após o abate, os pesos da gordura renal, pélvica e inguinal e da carcaça foram medidos e a carcaça dividida em músculos, ossos e aparas. Foram mensurados a área de olho de lombo e a espessura de gordura subcutânea sobre o músculo Longissimus na região da 12ª costela e foram obtidos bifes para a determinação da cor, força de cisalhamento e perdas por cocção e cozimento da carne. A levedura aumentou o rendimento de carcaça, mas não houve efeito dos tratamentos sobre o peso de carcaça, porção comestível e aparas. Os aditivos não influenciaram o pH da carcaça, a cor, a gordura intramuscular, a força de cisalhamento e as perdas por exsudação da carne. A suplementação, com levedura e com monensina em associação ou separadamente, não teve efeito importante sobre a qualidade da carcaça em novilhos terminados em confinamento.

Uncertainty measurement of chlorophenols and PCBs analyzed in aqueous media by SPME-GC-ECD.

Chlorophenols (CPs) and polychlorinated biphenyls (PCBs) are two of the most important groups of high-priority pollutants, due to their carcinogenicity, toxicity, and mutagenicity. This paper compares the utilization of two solid-phase microextraction (SPME) based extraction procedures, prior to gas chromatography with electron capture detection (GC-ECD), to determine PCBs (PCB 101 and PCB 153) and CPs (2,4,6-TCP and PCP) in aqueous media. Good linearity was observed with the SPME-GC-ECD method for the concentration range studied; detection limits ranged from 0.5 to 1.0 microg/L. Repeatability was between 13% and 31% for the lowest concentration and between 5.6% and 7.8% for the highest concentration. The uncertainty was determined by Thompson and bottom-up approaches. The identification of the compounds was confirmed by GC-MS. The developed procedure has the advantage of simplicity of sample treatment and avoids the use of potentially hazardous organic solvents and the clean-up or pre-concentration steps. Regarding PCBs, this procedure is simpler and faster, but the limits of detection are higher.

Perfil eletroforético do proteinograma sérico de eqüinos com obstrução experimental do cólon menor / Eletrophoretic profile of serum proteinogram of equines with small colon experimental obstruction

Avaliaram-se as alterações do proteinograma sérico de eqüinos submetidos à isquemia e reperfusão do cólon menor por distensão intraluminal. Foram utilizados 10 animais submetidos à laparotomia pelo flanco, em posição quadrupedal, para a indução de obstrução no cólon menor durante um período de quatro horas. Cinco animais foram instrumentados, mas sem distensão (grupo controle - G1). Em cinco outros animais, foi realizada isquemia mural por distensão do cólon menor via manguito inflado com 40mmHg (grupo distendido - G2). Foram colhidas amostras de sangue antes da intervenção cirúrgica (M1), com 4 horas da colocação do manguito (M2) e com 3 horas (M3) e 12 horas (M4) de reperfusão. Após centrifugação e fracionamento das amostras, as proteínas de fase aguda foram separadas por eletroforese em gel de poliacrilamida contendo SDS-PAGE, e suas concentrações determinadas por densitometria computadorizada. Foram encontradas 19 proteínas no fracionamento eletroforético, com peso molecular variando de 185.000 a 14.000 Daltons (Da). Os pesos moleculares encontrados, correspondentes às proteínas mais conhecidas, foram ceruloplasmina, 130.000 Da; proteína C-reativa, 122.000 Da; transferrina, 85.000 Da; α1-antitripsina, 61.000 Da; haptoglobina, 47.000 Da; e glicoproteína ácida, 40.000 Da. Os resultados mostram que proteínas de fase aguda se alteram após o trauma cirúrgico.

The changes in the serum proteinogram of equines submitted to ischemia and reperfusion of small colon by intraluminal distension were evaluated. Ten horses were submitted to standing laparotomy to induce a small colon obstruction for four hours. Five animals were instrumented without distention (control group - G1). In another five animals, mural ischemia was promoted placing a latex balloon distended with 40mmHg into the small colon (distended group - G2). Blood samples were collected before the surgical procedures (M1) and 4h after the balloon placement (M2) and 3h (M3) and 12h (M4) after reperfusion. After centrifugation and fractioning of the samples, the acute phase proteins were separated by SDS-PAGE and their concentrations were determined by computer densitometry. Nineteen proteins were separated by eletrophoresis and the molecular weights ranged from 185,000 to 14,000 Dalton (Da). The found molecular weights, related to the most known proteins, were: ceruloplasmin, 130,000 Da; C-reactive protein, 122,000 Da; transferrin, 85,000 Da; α1-antitripsin, 61,000 Da; haptoglobin, 47,000 Da; and acid glycoprotein, 40,000 Da. The results showed that acute phase proteins changed after surgical injury.

Leucograma e teores plasmáticos de proteínas de fase aguda de eqüinos portadores de abdômen agudo e submetidos à laparotomia / Leukogram and plasma acute phase protein concentrations in horses with acute abdomen submitted to treatment by laparotomy

Foram examinados 20 eqüinos adultos portadores de abdômen agudo e submetidos à laparotomia. Dez recuperaram-se sem intercorrência pós-operatória (G1) e 10 foram a óbito sete a 10 dias após a cirurgia, com sinais de choque séptico (G2). Avaliaram-se temperatura retal, freqüências cardíaca e respiratória, tempo de preenchimento capilar e teores plasmáticos das proteínas de fase aguda - fibrinogênio, ceruloplasmina, proteína C-reativa, antitripsina, haptoglobina e glicoproteína ácida -, antes e até sete dias após a laparotomia. As leucometrias às 72h e no sétimo dia pós-operatório dos eqüinos que foram a óbito foram, respectivamente, 34,6 por cento e 57,1 por cento, mais altas que a dos animais curados. Os maiores valores de proteína de fase aguda ocorreram no sétimo dia após a cirurgia; os percentuais de elevação de fibrinogênio, antitripsina, glicoproteina ácida, proteína C-reativa, ceruloplasmina e haptoglobina de eqüinos do G2 em relação ao G1 foram 46,8 por cento, 67,9 por cento, 91,9 por cento, 112,2 por cento, 126,9 por cento e 186,2 por cento, respectivamente.

Twenty adult horses with acute abdomen were examined and submitted to treatment by laparotomy; ten had no postoperative complication (group 1), and ten showed septic shock symptom and died from seven to ten days after surgery (group 2). Body temperature, heart and respiratory rates, filling capillary time, and plasma acute phase protein concentrations - fibrinogen, ceruloplasmin, C-reactive protein, antitrypsin, haptoglobin, and acid glycoprotein - were evaluated before laparotomy and until seven days after surgery. White blood cell counts at 72h and seven days after surgery in group 2 animals were, respectively, 34.6 percent and 57.1 percent, and were higher than those measured in group 1 horses. The highest values of acute phase proteins occurred on the seventh day after surgery. The increase percentages of fibrinogen, antitrypsin, acid glycoprotein, C-reactive protein, ceruloplasmin, and haptoglobin plasma concentrations in group 2 animals in comparison to group 1 animals were 46.8 percent, 67.9 percent, 91.9 percent, 112.2 percent, 126.9 percent, and 186.2 percent, respectively.