Synthesis and glycosidase inhibitory profiles of functionalised morpholines and oxazepanes

In this work libraries of morpholines and oxazepanes have been prepared via the reductive amination reaction between dialdehydes, derived from carbohydrates, and a range of amines. In this way, functionalised morpholines and oxazepanes have been prepared that include N-alkylated derivatives, disaccharide analogues, and ester containing derivatives. The abilities of these functionalised morpholines and oxazepanes to inhibit a broad panel of glycosidase enzymes, that are associated with a range of diseases, have been probed and in this way new inhibitors of a range of glycosidases, but particularly β-d-galactosidase derived from Bovine kidney, have been discovered. N-Alkyl morpholines demonstrated the best inhibition profiles for this enzyme and derivatives (15a)–(15d) acted as non-competitive inhibitors with IC50 values of 55.1–88.6 μM. Within this study, some preliminary structure–activity relationships are proposed, and it is demonstrated that N-substituted morpholines display better inhibitory profiles for the enzymes analysed than any of the N-substituted oxazepanes.

Sustained polymeric delivery of gene silencing antisense ODNs, siRNA, DNAzymes and ribozymes: in vitro and in vivo studies

Small interfering RNA (siRNA), antisense oligonucleotides (ODNs), ribozymes and DNAzymes have emerged as sequence-specific inhibitors of gene expression that may have therapeutic potential in the treatment of a wide range of diseases. Due to their rapid degradation in vivo, the efficacy of naked gene silencing nucleic acids is relatively short lived. The entrapment of these nucleic acids within biodegradable sustained-release delivery systems may improve their stability and reduce the doses required for efficacy. In this study, we have evaluated the potential in vitro and in vivo use of biodegradable poly (d,l-lactide-co-glycolide) copolymer (PLGA) microspheres as sustained delivery devices for ODNs, ribozyme, siRNA and DNA enzymes. In addition, we investigated the release of ODN conjugates bearing 5′-end lipophilic groups. The in vitro sustained release profiles of microsphere-entrapped nucleic acids were dependent on variables such as the type of nucleic acid used, the nature of the lipophilic group, and whether the nucleic acid used was single or double stranded. For in vivo studies, whole body autoradiography was used to monitor the bio-distribution of either free tritium-labelled ODN or that entrapped within PLGA microspheres following subcutaneous administration in Balb-c mice. The majority of the radioactivity associated with free ODN was eliminated within 24 h whereas polymer-released ODN persisted in organs and at the site of administration even after seven days post-administration. Polymer microsphere released ODN exhibited a similar tissue and cellular tropism to the free ODN. Micro-autoradiography analyses of the liver and kidneys showed similar bio-distribution for polymer-released and free ODNs with the majority of radioactivity being concentrated in the proximal convoluted tubules of the kidney and in the Kupffer cells of the liver. These findings suggest that biodegradable PLGA microspheres offer a method for improving the in vivo sustained delivery of gene silencing nucleic acids, and hence are worthy of further investigation as delivery systems for these macromolecules.

Hippurate: the natural history of a mammalian-microbial co-metabolite

Hippurate, the glycine conjugate of benzoic acid, is a normal constituent of the endogenous urinary metabolite profile and has long been associated with the microbial degradation of certain dietary components, hepatic function and toluene exposure, and is also commonly used as a measure of renal clearance. Here we discuss the potential relevance of hippurate excretion with regards to normal endogenous metabolism and trends in excretion relating to gender, age, and the intestinal microbiota. Additionally, the significance of hippurate excretion with regards to disease states including obesity, diabetes, gastrointestinal diseases, impaired renal function, psychological disorders and autism, as well as toxicity and parasitic infection, are considered.

alpha-Hydroxynitrile lyase protein from Xylella fastidiosa: Cloning, expression, and characterization

Xylella fastidiosa is a xylem-restricted plant pathogen that causes a range of diseases in several and important crops. Through comparative genomic sequence analysis many genes were identified and, among them, several potentially involved in plant-pathogen interaction. The experimental determination of the primary sequence of some markedly expressed proteins for X fastidiosa and the comparison with the nucleic acids sequence of genome identified one of them as being SCJ21.16 (XFa0032) gene product. The comparative analysis of this protein against SWISSPROT database, in special, resulted in similarity with a-hydroxynitrile lyase enzyme (HNL) from Arabidopsis thaliana, causing interest for being one of the most abundant proteins both in the whole cell extract as well as in the extracellular protein fraction. It is known that HNL enzyme are involved in a process termed ""cyanogenesis"", which catalyzes the dissociation of alpha-hydroxinitrile into carbonyle and HCN when plant tissue is damaged. Although the complete genome sequences of X.fastidiosa are available and the cyanogenesis process is well known, the biological role of this protein in this organism is not yet functionally characterized. In this study we presented the cloning, expression, characterization of recombinant HNL from X fastidiosa, and its probable function in the cellular metabolism. The successful cloning and heterologous expression in Escherichia coli resulted in a satisfactory amount of the recombinant HNL expressed in a soluble, and active form giving convenient access to pure enzyme for biochemical and structural studies. Finally, our results confirmed that the product of the gene XFa0032 can be positively assigned as FAD-independent HNLs. (C) 2009 Elsevier Ltd. All rights reserved.

Serosurvey for antibodies against Brucella abortus and Leptospira interrogans in pampas deer from Brazil

A survey for antibodies against Brucella abortus, and Leptospira interrogans was conducted on 17 pampas deer (Ozotocerus bezoarticus) from Pantanal Matogrossense (State of Mato Grosso do Sul, Brazil) and on 24 pampas deer from Parque Nacional de Emas (State of Goias, Brazil). Antibodies against B. abortus were detected by plate ag glutination, rose Bengal, and complement fixation tests, antibodies against Leptospira interrogans were detected by the microscopic agglutination test. All sera were negative for B. abortus antibodies and all deer sera from Parque Nacional de Emas were negative for L. interrogans antibodies. Four (24%) of 17 sera from Pantanal Matogrossense were positive for L. interrogans serovar (n = 2) hardjo, wolffi (n = 1) and mini (n = 1). While these diseases do not appear to be of major importance to the health status of Pampas deer, it appears that deer are reservoir for leptospirosis in one of the study areas.

Qualidade de vida de pessoas com úlcera venosa: associação dos aspectos sociodemográficos, de saúde, assistência e clínicos da lesão

Venous ulcer (VU) is a lower limbs injury resulting from inadequate return of venous blood in feet or legs. Although it is not a deadly disease, it causes chronic wounds, which seriously undermine patients´ quality of life (QOL) and sometimes leads to drastic family, social, economic and psychological changes. In this sense, there are several aspects that may influence the venous ulcers patients´ QOL. The study´s objective aimed on the association of socio-demographic and health, health care and clinical injury on UV patients‟ QOL. Analytical studies, which consider the complexity of factors involved in changes in UV patients‟ QOL has a cross-sectional and quantitative approach. The HUOL Ethics Committee approved this project (n.279/09). The collection of data lasted a period of 3 months in 2010 and it took place at the clinic of Angiology at Hospital Universitário Onofre Lopes (HUOL). The data sample consisted of 60 patients treated by UV angiologists in the HUOL Surgical Clinic. The results were analyzed with SPSS 15.0 by descriptive and inferential statistics. The study was based on UV patients that were predominantly female, average age of 61.4 years, that had low education level and low family income, with occupations requiring long periods of standing or sitting, but mostly retired, unemployed or laid off due to the disease and/or due to chronic diseases associated with the UV. The study took also into consideration patients that used inappropriate products, that were improperly treated by a professional caregiver, that lacked of adequate guidance and compression therapy, that performed no lifting of the lower limbs and regular exercise, that the time of injury were greater than or equal to six months, that were missing specific laboratory tests. The study‟s reference were on recurrent lesions, medium to large lesions area, bed of the lesion (injuries) with fibrin and/or necrosis, with amount of exudate with medium to large, odorless and no signs of infection, with tissue loss between 1st and 2nd degree, without collecting swab or biopsy and with pain. In general, QOL of researched individuals were considered low, the maximum score was 69 points, which the areas that were mostly influenced were the total scores of QOL functional capacity (0.021), emotional (0.000) and social functioning (0.080). Of the 60 individuals, 53.3% had scores between 40 and 69 points in SF-36, and they had the best scores in sociodemographic and health variables (ρ = 0.049). In respect to the assistance and injury characteristics, patients who scored between 40 and 69 points in SF-36 had better scores on these characteristics. By combining the socio-demographic variables, health, and handling characteristics of the injury, we observed a significant difference (ρ = 0.032) when linking them with the QOL total scores. When analyzing separately the domains of the SF-36 scores on the quality of life, we find that the areas that showed statistical significance were functional ability (ρ = 0.035), appearance (ρ = 0.019), emotional (ρ = 0.000), and mental health (ρ = 0.050). Among the socio-demographic characteristics studied, gender and marital status contributed more to the reduction of QOL and among the variables of assistance and the injury, orientation, reference and area of UV contributed the most. By analyzing these five variables all together in accordance with the overall score obtained in the quality of life, we found a significant correlation (ρ = 0.002); with 6.23 times more chances of patients have better QOL in the presence of these five positive factors. By conducting the Mann Whitney U test between all the five demographic variables, health, and clinical care, we found that this combination also proved to be significant (ρ = 0.006). Therefore, patients with these five variables positive tend to have a better QOL. Based on these results, we reject the null hypothesis (H0) and accept the alternative hypothesis (H1) proposed in this study because we noted that the QOL of patients with UV is associated with sociodemographic and health, health care and clinical aspects of the injury

First Identification of Canine Distemper Virus in Hoary Fox (Lycalopex vetulus): Pathologic Aspects and Virus Phylogeny

Canine distemper virus (CDV) has been reported in several wild animal species, but there have been no reports of CDV in hoary fox (Lycalopex vetulus). This paper characterizes the first case of natural CDV infection in hoary fox, including the clinical and pathologic aspects of the disease as well as the viral strain phylogeny.

Emerging infectious diseases in cetaceans worldwide and the possible role of environmental stressors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Hemoglobina Köln diagnosticada em programa de triagem neonatal em São José do Rio Preto, SP

Alterações genéticas em que a mutação de aminoácidos nas globinas afeta a estrutura da molécula tornando-a instável são classificadas como hemoglobinas instáveis. Devido à grande diversidade dos pontos de mutações por substituições e deleções de aminoácidos, as formas de instabilização se apresentam muito variadas. A hemoglobina Köln é a variante instável descrita com maior freqüência na literatura e a terceira descoberta no Brasil, as outras são Hb Niterói e Hb Hasharon. Anemia moderada, icterícia e presença de urina escura caracterizam as manifestações clínicas da Hb Köln. em programa de triagem neonatal identificamos uma criança com suspeita de heterozigose para hemoglobina Köln, confirmada por procedimentos eletroforéticos e HPLC. Avaliações por diferentes metodologias laboratoriais e estudo familiar auxiliam no diagnóstico precoce, possibilitando minimizar os sintomas decorrentes da hemoglobina anormal e a realização do aconselhamento genético e educacional destas alterações hereditárias.

alpha-Hydroxynitrile lyase protein from Xylella fastidiosa: Cloning, expression, and characterization

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Molecular and Serologic Detection of Ehrlichia spp. in Endangered Brazilian Wild Captive Felids

Ehrlichiosis, an emergent tickborne disease that affects both humans and animals, may represent a threat to the survival and preservation of wild felids in Brazil There are few studies of ehrlichiosis in wild felids in Brazil, but Ehrlichia spp are present in domestic cats Antibodies to Ehrlichia canis have been reported in a puma (Puma concolor) In this study we assessed the presence of these hemoparasites in the blood of Brazilian wild captive felids of the 72 animals tested, 5 (7%) were seropositive for the E cams antigen, and L1 (15%) were positive for E emirs DNA sequences We also performed sequence alignment to establish the identity of the parasite species infecting these animals using 16S rRNA and omp-1 genes Sequences based on 16S rRNA were similar to those found in dogs and cats from Thailand, Brazil, China, and Taiwan and with E canis obtained from a single individual (human) in Venezuela Ehrlichia sp sequence from sampled felines based on omp-1. gene was similar to the p28 and p30 multigene family of E canis To our knowledge, this is the first study of molecular detection of Ehrlichia sp in Brazilian wild feline species

Ocorrência de enteroparasitos em centros de educação infantil no Município de Patos de Minas, MG, Brasil

Considering the high prevalence of the parasitic diseases in Brazil and its close relationship with the socio-economic and cultural levels of the population, aimed to establish the occurrence of enteroparasites in children of six Centers of Education Child (CEIs) of the municipality of Patos of Minas (MG). Were collected three samples of faeces of 161 children from zero to six years of age. Were used the Lutz and Baermann-Moraes' methods, to the detection of cysts of protozoa, eggs and larvae of helminths. Coproparasitological analyses showed positivity of 73%. The most frequent helminths were: Ascaris lumbricoides (50%), hookworm (22%) and Enterobius vermicularis (0.6%). Among the protozoa highlighted were: Entamoeba histolytica/E. dispar (22%) and Giardia lamblia (32%). The high occurrence of enteroparasites evidenced the need for an effective health policy in the CEIs of the municipality of Patos of Minas to the fight against intestinal parasites.

Molecular detection of Cytauxzoon spp. in asymptomatic Brazilian wild captive felids

Cytauxzoon. spp. DNA was detected for the first time in blood samples from asymptomatic Brazilian wild captive felids. In 2606, 72 EDTA blood samples from seven wild felids species: Puma concolor (pinna), Leopardus pardalis (ocelot), Puma yagouaroundi (jaguarundi), Leopardus wiedii (margay), Leopardus tigrinus (little spotted cat), Oncifelis colocolo (pampas cat) and Panthera. onca. (jaguar) were analyzed using polymerase chain reaction to amplify the 18S rRNA gene segment in order to verify the presence of Cytauxzoon spp. DNA. Nine samples were positive: six ocelots, two pumas, and one jaguar. In Brazil, wild felids may be natural reservoirs for Cytauxzoon spp.

The glandular odontogenic cyst: a case report.

A case of a glandular odontogenic cyst is reported. All clinical, radiographic, and histopathologic features are discussed and compared with the descriptions in the literature already published about this new lesion.