Search

Chlamydiae are a large group of obligate endosymbionts of eukaryotes that includes the Chlamydiaceae family, comprising several animal pathogens. Among Chlamydiaceae, Chlamydia trachomatis causes widespread ocular and urogenital infections in humans. Like many bacterial pathogens, all Chlamydiae manipulate host cells by injecting them with type III secretion effector proteins. We previously characterized the C. trachomatis effector CteG, which localizes at the host cell Golgi and plasma membrane during distinct phases of the chlamydial infectious cycle. Here, we show that CteG is a Chlamydiaceae-specific effector with over 60 homologs phylogenetically categorized into two distinct clades (CteG I and CteG II) and exhibiting several inparalogs and outparalogs. Notably, cteG I homologs are syntenic to C. trachomatis cteG, whereas cteG II homologs are syntenic among themselves but not with C. trachomatis cteG. This indicates a complex evolution of cteG homologs, which is unique among C. trachomatis effectors, marked by numerous events of gene duplication and loss. Despite relatively modest sequence conservation, nearly all tested CteG I and CteG II proteins were identified as type III secretion substrates using Yersinia as a heterologous bacterial host. Moreover, most of the type III secreted CteG I and CteG II homologs were delivered by C. trachomatis into host cells, where they localized at the Golgi region and cell periphery. Overall, this provided insights into the evolution of bacterial effectors and revealed a Chlamydiaceae family of type III secreted proteins that underwent substantial divergence during evolution while conserving the capacity to localize at specific host cell compartments. [ABSTRACT FROM AUTHOR]

Neutropenia and neutrophil dysfunction in glycogen storage disease type 1b (GSD1b) and severe congenital neutropenia type 4 (SCN4), associated with deficiencies of the glucose-6-phosphate transporter (G6PT/SLC37A4) and the phosphatase G6PC3, respectively, are the result of the accumulation of 1,5-anhydroglucitol-6-phosphate in neutrophils. This is an inhibitor of hexokinase made from 1,5-anhydroglucitol (1,5-AG), an abundant polyol in blood. 1,5-AG is presumed to be reabsorbed in the kidney by a sodium-dependent-transporter of uncertain identity, possibly SGLT4/SLC5A9 or SGLT5/SLC5A10. Lowering blood 1,5-AG with an SGLT2-inhibitor greatly improved neutrophil counts and function in G6PC3-deficient and GSD1b patients. Yet, this effect is most likely mediated indirectly, through the inhibition of the renal 1,5-AG transporter by glucose, when its concentration rises in the renal tubule following inhibition of SGLT2. To identify the 1,5-AG transporter, both human and mouse SGLT4 and SGLT5 were expressed in HEK293T cells and transport measurements were performed with radiolabelled compounds. We found that SGLT5 is a better carrier for 1,5-AG than for mannose, while the opposite is true for human SGLT4. Heterozygous variants in SGLT5, associated with a low level of blood 1,5-AG in humans cause a 50–100% reduction in 1,5-AG transport activity tested in model cell lines, indicating that SGLT5 is the predominant kidney 1,5-AG transporter. These and other findings led to the conclusion that (1) SGLT5 is the main renal transporter of 1,5-AG; (2) frequent heterozygous mutations (allelic frequency > 1%) in SGLT5 lower blood 1,5-AG, favourably influencing neutropenia in G6PC3 or G6PT deficiency; (3) the effect of SGLT2-inhibitors on blood 1,5-AG level is largely indirect; (4) specific SGLT5-inhibitors would be more efficient to treat these neutropenias than SGLT2-inhibitors. [ABSTRACT FROM AUTHOR]

In swallowing, the hyoid bone moves up and forward in response to the activation of suprahyoid muscles, opening the upper esophageal sphincter and aiding the airway protection mechanism. This displacement measure has been analyzed with ultrasound images because this method does not expose the patient to radiation, has a good cost–benefit ratio, and is safe for the patient. However, there is no consensus on the reliability of this ultrasound measure. The objective of this study was to analyze the reliability of measuring hyoid bone displacement amplitude in swallowing with ultrasound. The systematic review encompassed five databases (MEDLINE, Scopus, EMBASE, Web of Science, Cochrane Library) and gray literature. There was no limitation of language or year of publication. The search/selection/extraction methodology was conducted by two authors blindly and independently, and differences were solved by a third rater. Three studies met the eligibility criteria: two of them analyzed the reliability in non-dysphagic populations and the other, in dysphagic patients. The transducer was positioned in the submandibular region in all studies. The authors were not clear about the training time to acquire and analyze ultrasound images. The meta-analysis had an interrater reliability of 0.858 (95% CI: 0.744–0.924) and intrarater reliability of 0.968 (95% CI: 0.903–0.990). There was, however, heterogeneity of p = 0.005 for intrarater reliability. Despite good reliability, the heterogeneity reinforces the importance of training and protocol standardization for image acquisition and analysis. [ABSTRACT FROM AUTHOR]

Transaminases play key roles in central metabolism, transferring the amino group from a donor substrate to an acceptor. These enzymes can often act, with low efficiency, on compounds different from the preferred substrates. To understand what might have shaped the substrate specificity of this class of enzymes, we examined the reactivity of six human cytosolic transaminases towards amino acids whose main degradative pathways do not include any transamination. We also tested whether sugars and sugar phosphates could serve as alternative amino group acceptors for these cytosolic enzymes. Each of the six aminotransferases reacted appreciably with at least three of the alternative amino acid substrates in vitro, albeit at usually feeble rates. Reactions with L-Thr, L-Arg, L-Lys and L-Asn were consistently very slow—a bias explained in part by the structural differences between these amino acids and the preferred substrates of the transaminases. On the other hand, L-His and L-Trp reacted more efficiently, particularly with GTK (glutamine transaminase K; also known as KYAT1). This points towards a role of GTK in the salvage of L-Trp (in cooperation with ω-amidase and possibly with the cytosolic malate dehydrogenase, MDH1, which efficiently reduced the product of L-Trp transamination). Finally, the transaminases were extremely ineffective at utilizing sugars and sugar derivatives, with the exception of the glycolytic intermediate dihydroxyacetone phosphate, which was slowly but appreciably transaminated by some of the enzymes to yield serinol phosphate. Evidence for the formation of this compound in a human cell line was also obtained. We discuss the biological and evolutionary implications of our results. [ABSTRACT FROM AUTHOR]

Summary: Hip fracture incidence rates in three representative geographic areas in Brazil over a period of 2 years (2010–2012) were assessed for the first time. Estimated incidence rates varied regionally, and markedly differed from those previously reported. Thus, national guidelines as well as FRAX Brazil should be revised in light of this new data. Purpose: To determine the annual incidence of hip fractures in individuals aged 50 years and over, living in 3 cities located in different regions of the country. To investigate the age, gender, and regional differences in fracture rates. Based on the obtained data, to estimate the national incidence of hip fractures resulting from osteoporosis, in order to improve prevention strategies. Methods: Retrospective, observational study including all patients aged ≥ 50 years admitted in hospitals because of a hip fracture in three cities (Belem, Joinville, and Vitoria) from representative geographic areas in Brazil from 2010 to 2012. Data were obtained from medical records in those cities. We analyzed incidence rates (crude and age- and gender-standardized rates) for hip fractures. Results: There were 1025 (310 in men and 715 in women) hip fractures in the over 50-year-old merged population from the three cities. The crude incidence rate for hip fracture was 103.3/100,000 (95% confidence interval [CI = 97.0; 109.7), in men 77.4/100,000 (95% CI = 68.8; 86.0), and in women 125.2/100,000 (95% CI = 116.0; 134.4). Incidence standardized for age and gender was 105.9 cases per 100,000 persons per year (95% CI = 99.4; 112.4); 78.5 cases per 100,000 (95% CI = 69.8; 87.3) in men and 130.6 cases 100,000 in women (95% CI = 121.0, 140.2) per year. Belem, located in the equatorial region (latitude 1° 27′ S), had significantly lower crude and age-adjusted incidence than Joinville (latitude 26° 18′ S) and Vitoria (latitude 20° 19′ S), which were no different from each other. The incidence of fractures increased exponentially with age, and women had about twice the risk of fractures than men. Conclusions: Hip fracture mainly affects elderly women and presents great variability in incidence between the different regions in Brazil. The incidence of hip fractures in Brazil differed markedly from that reported previously, so that national guidelines and the FRAX model for Brazil should be revised. [ABSTRACT FROM AUTHOR]

This work aimed to develop an edible coating based on Mexican lime (Citrus aurantifolia Swingle) pectic extract and essential oil on Haden mango (Mangifera indica L.) to extend its shelf life. Mango cubes were coated by immersion in a lime pectic extract (1% pectin w/v), lime essential oil (0.05% v/v), and glycerol (0.7% v/v) solution for 2, 5, and 10 min. Subsequently, coated and uncoated (control) test samples were stored for 21 days, and physical–chemical and microbiological analyses were performed every 3 days. The results showed no significant differences for total soluble solids, pH, and firmness. On the sixth day, bacterial growth was significantly lower in coated mangos than in the control (log 6.08 ± 0.49 and 7.63 ± 0.20 UFC g−1, respectively). The application of the edible coating extended the shelf life of minimally processed mangos by 3 days, delaying physical and chemical changes as well as bacterial growth. [ABSTRACT FROM AUTHOR]

Phosphoglucomutase 3 (PGM3) deficiency is a rare congenital disorder of glycosylation. Most of patients with autosomal recessive hypomorphic mutations in PGM3 encoding for phosphoglucomutase 3 present with eczema, skin and lung infections, elevated serum IgE, as well as neurological and skeletal features. A few PGM3-deficient patients suffer from a more severe disease with nearly absent T cells and severe skeletal dysplasia. We performed targeted next-generation sequencing on two kindred to identify the underlying genetic etiology of a severe combined immunodeficiency with developmental defect. We report here two novel homozygous missense variants (p.Gly359Asp and p.Met423Thr) in PGM3 identified in three patients from two unrelated kindreds with severe combined immunodeficiency, neurological impairment, and skeletal dysplasia. Both variants segregated with the disease in the two families. They were predicted to be deleterious by in silico analysis. PGM3 enzymatic activity was found to be severely impaired in primary fibroblasts and Epstein–Barr virus immortalized B cells from the kindred carrying the p.Met423Thr variant. Our findings support the pathogenicity of these two novel variants in severe PGM3 deficiency. [ABSTRACT FROM AUTHOR]

Peritonitis due to gram-negative bacilli (GNB), particularly nonfermenting GNB (NF-GNB), is a serious complication of peritoneal dialysis with a low resolution rate. Beyond the patient's condition, microbiological properties such as antimicrobial resistance, biofilm production and other virulence factors can explain the poor outcomes. This study aimed to evaluate the influence of patient condition, microbiological characteristics, including biofilm production, and treatment on peritonitis outcome. We reviewed the records of 62 index episodes caused by NF-GNB that occurred between 1997 and 2015 in our center. The etiologies were species of Pseudomonas (51.6%), Acinetobacter (32.2%), and other NF-GNB (16.1%). There was a high (72.9%) proportion of biofilm producer lineages. The in vitro susceptibility rate of Pseudomonas spp. to amikacin, ciprofloxacin, and ceftazidime was significantly greater than that of Acinetobacter spp. and other species; however, there was a similar low resolution rate (< 45%) among the episodes attributable to them. Preexisting exit-site infection was independently associated with nonresolution. No other factor, including biofilm production, was associated with the outcome. The higher in vitro susceptibility of Pseudomonas compared to other NF-GNB that presented a similar resolution rate suggests that bacterial virulence factors such as biofilms can act in concert, thereby worsening the outcome. [ABSTRACT FROM AUTHOR]

Blastocystis sp. is an intestinal protist parasite commonly found in the feces of humans and animals worldwide. Blastocystis exhibits extensive genetic diversity and has been identified in humans and a variety of animals including other mammals and birds. Blastocystis subtypes do not exhibit strict host specificity which raises the possibility of zoonotic transmission through either direct contact or fecal contamination of food or water. However, reports detailing the subtypes and prevalence of Blastocystis in avian species are limited. Therefore, this study investigated the presence of Blastocystis in chickens by molecular characterization of the small subunit rRNA (SSU rRNA) gene. Fecal samples from 130 chickens were collected from local markets in Uberlândia and Belo Horizonte in the state of Minas Gerais, Brazil. To detect and identify subtypes of Blastocystis, a next-generation amplicon sequencing protocol was used. Forty-four of the 130 (33.8%) chickens examined were positive for Blastocystis. Blastocystis subtypes ST6 (23/130; 17.7%), ST7 (43/130; 33.1%), ST10 (1/130; 0.8%), ST14 (5/130; 3.8%), ST25 (1/130; 0.8%), and a novel subtype (ST29) (2/130; 1.5%) were observed. A nanopore sequencing strategy was used to obtain the near full-length SSU rRNA gene nucleotide sequence and validate novel subtype ST29. Mixed infections containing multiple subtypes were common and identified in 63.6% of Blastocystis-positive chickens. All positive samples contained one or both potentially zoonotic subtypes ST6 and ST7. The prevalence of Blastocystis in chickens was high, and molecular characterization mostly identified subtypes previously found in humans. Thus, chickens may be a source of human infection and environmental contamination. [ABSTRACT FROM AUTHOR]

Root-knot nematodes (RKN), Meloidogyne spp., are important crop pests that cause severe losses in crop production worldwide, reducing both productivity and crop quality. Meloidogyne chitwoodi Golden, O'Bannon, Santo & Finley, 1980 is considered a quarantine organism by the European and Mediterranean Plant Protection Organization (EPPO) causing damage in tomato and potato crops. The development of nonchemical and sustainable management strategies to reduce nematode damage is crucial. The resistance of Solanum linnaeanum Hepper & P.-M.L. Jaeger and S. sisymbriifolium Lamarck cv. Sis 6001 to M. chitwoodi was evaluated based on gall index (GI), the Bridge & Page (1980) rating chart and reproduction factor (RF). Both plant species were resistant to M. chitwoodi. Solanum linnaeanum had an average of 519 small root swellings/plant, with 45% adult nematodes inside the roots, all males. Solanum sisymbriifolium had GI ≤ 2 and RF ≤ 1 with a high percentage (69%) of nematodes inside the roots that did not develop beyond the sexually undifferentiated second-stage. The use of S. linnaeanum as a new source of resistance is a good alternative for the control of RKN in the quest to develop nonchemical and sustainable management strategies to protect crops. [ABSTRACT FROM AUTHOR]

Purpose: Peritonitis is a serious complication of peritoneal dialysis and coagulase-negative Staphylococcus (CNS) is the most frequent cause of peritoneal dialysis (PD)–infections in many centers. This study aimed to investigate the molecular epidemiology of CNS isolated from PD-peritonitis in a Brazilian single center, focusing on the genetic determinants conferring methicillin resistance. Methods: Bacterial strains were isolated from peritoneal fluid of patients presenting PD-peritonitis, identified by phenotypic and molecular methods, and those identified as CNS were submitted to mecA detection, SCCmec, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results: Over the 18-year period of this study (1995–2011), a total of 878 peritonitis episodes were diagnosed in this unit, 115 were caused by coagulase-negative staphylococci of which 72 by Staphylococcus epidermidis. mecA gene was detected in 55 CNS (47.8%), more frequently on the more recent years. SCCmec type III was the most frequent cassette, followed by SCCmec type IV and SCCmec type II. A diverstity of pulsotypes was observed among the S. epidermidis isolates, but five clusters (based on the 80% cutoff) were identified. Diversified sequence types (ST02, ST05, ST06, ST09, ST23, ST59 and ST371) were detected. Conclusions: Detection of SCCmec type III among coagulase-negative Staphylococcus underscores the role of hospital environments as potential source of methicillin-resistant Staphylococcus causing peritonitis in PD patients. [ABSTRACT FROM AUTHOR]

Babesia is tick-transmitted protozoan parasites that infect mammalian hosts and have a major impact on farm and pet health-associated costs worldwide. This study aimed to test the prevalence of Babesia spp. infection in a small cohort of dogs at a veterinary hospital and to perform molecular characterization of the Babesia species causing the infection. For the PCR assay, 5 mL of blood was collected by venipuncture of the cephalic or radial veins in 300 dogs of different ages, sex, and breeds, which were presented to the veterinary hospital of the Federal University of Uberlândia between March 2015 and April 2016. In addition, a drop of blood was collected from the marginal blood vessels of the ear of dogs included in this study. Ninety-two (30.67%) were positive for Babesia spp., as determined by microscopic observation of the blood smear, revealing the presence of intra-erythrocyte merozoites. For molecular characterization by PCR, 17 samples were chosen from dogs who were tested positive for Babesia spp. by blood smears. Among them, B. vogeli was found to infect all 17 dogs, as determined by 99–100% sequence identity (closest GenBank match KT246307) using primers PIRO A/PIRO B. Our results indicate that the species observed in these dogs was B. vogeli. [ABSTRACT FROM AUTHOR]

Species of Hepatozoon are parasites frequently recorded in snakes. The species identification of this genus was based mostly on the gametocyte morphology and morphometric calculations. For more reliable results, molecular characterization, an initial step for the correct identification of Hepatozoon species, has been used. This study aimed to determine the prevalence and identification of Hepatozoon species in captive snakes from Brazil. To that end, morphological, morphometric, and molecular data were obtained. A total of 157 snakes; 128 venomous (Crotalus durissus) and 29 non-venomous (Epicrates crassus and Boa constrictor) were screened for Hepatozoon blood parasites. Using light microscopy, 20 (12.78%) snakes were found positive for Hepatozoon spp., of which 6/29 (20.7%) were non-venomous and 14/128 (10.9%) were venomous; all with low parasitemia. Polymerase chain reaction (PCR), performed with the primers HepF300/Hep900, confirmed all 20 (100%) samples positive for hemogregarines. Species of Hepatozoon were identified from eight sequenced samples. Two previously described species, Hepatozoon cuestensis and Hepatozoon musa, were identified. The present study is the first to report H. musa within the snake hosts E. crassus and C. durrisus. In addition, a potentially new Hepatozoon species from B. constrictor was identified. [ABSTRACT FROM AUTHOR]

Herpes simplex virus 2 (HSV-2) is a very rare cause of central nervous system (CNS) infections. We report a case of a young woman with a left middle cerebral artery (MCA) ischemic stroke. The patient had history of HIV-1 infection, with periods of therapeutic non-compliance. Initial computed tomography (CT) imaging studies showed stenosis of the M1 segment of the left MCA, and magnetic resonance imaging (MRI) confirmed infarction of the MCA territory. Serial transcranial Doppler ultrasound revealed progressive occlusion of the MCA and stenosis of the left anterior cerebral artery. Systemic investigation for other causes of stroke was normal. Lumbar puncture revealed a mildly inflammatory cerebrospinal fluid, and HSV-2 DNA was identified by PCR, with a positive viral load in favor of active replication. No other viral or microbiological infections were identified. MRI angiography confirmed a vasculitic process involving the left carotid artery, and a HSV-2 vasculitis diagnosis was assumed. The patient started acyclovir with improvement of clinical features and imaging abnormalities. In the HIV-infected patient, stroke is a multifactorial common cause of morbidity. The physician should take into account a broad differential diagnosis including rare causes and atypical presentations of common etiologies, including HSV-1 and HSV-2 CNS infection. [ABSTRACT FROM AUTHOR]

The present work aimed at the construction and characterization of a multilayer coating based on κ-carrageenan and quercetin-loaded lecithin/chitosan nanoparticles (Np) by the layer-by-layer technique and the evaluation of its antioxidant capacity and potential cytotoxicity in vitro. The multilayered coating was successfully self-assembled, as confirmed by UV-Vis spectroscopy, contact angle, atomic force microscopy (AFM), and scanning electron microscopy (SEM). Multilayered coatings showed to have antioxidant capacity, with a DPPH• radical scavenging activity of 31.32 ± 3.13% and a result of the FRAP assay of 799.41 ± 95.39 μM of ferrous ion (Fe2+) equivalent. These coatings were also shown to be devoid of cell toxicity, as evaluated by determination of nitric oxide production and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. The alveolar macrophages culture was tested in the presence of the κ-carrageenan/quercetin-Np multilayer coating and showed a cell viability of 91.3 ± 9.6%. These results suggest that this multilayered coating is adequate for surfaces modification in view of biomedical and food industry applications. [ABSTRACT FROM AUTHOR]

The aim of this research was to evaluate the efficiency of liquid smoking and chitosan coating on the shelf life of Nile tilapia ( Oreochromis niloticus) fillets. Fillets without liquid smoked and chitosan coating (control), liquid smoked fillets (LS), and liquid smoked and chitosan coated fillets (LSCh) were stored at 4 ± 1 °C for 30 days. The physicochemical (pH, moisture content, water activity-a, color, texture, total volatile bases nitrogen-TVB-N and thiobarbituric acid reactive substances-TBARS) and microbiological analyses (mesophilic and psychrotrophic counts) and the electrophoresis profile of samples were carried out. Physicochemical parameters, such as TVB-N and TBARS, were reduced in the tilapia fillets with liquid smoking. The presence of the coating of chitosan was effective for the control of the microorganisms during storage. This work showed that the addition of a chitosan coating in liquid-smoked fillets further enhanced the effect of preservation. [ABSTRACT FROM AUTHOR]

A total of 85 fecal samples from captive birds collected from October 2013 to September 2014 in Uberlândia and Belo Horizonte in the state of Minas Gerais (Brazil) were evaluated for the presence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia by PCR. Of these, three birds were found positive for E. bieneusi (3.5%), two for Cryptosporidium (2.3%), and one for Giardia (1.2%). Two genotypes of E. bieneusi were detected by nucleotide sequence analysis of the ITS region, genotypes D and Peru 6 in a swan goose and in two rock pigeons, respectively. For Cryptosporidium and Giardia, nucleotide sequence analysis of the SSU rRNA identified Cryptosporidium baileyi and Duck genotype in a swan goose and a mandarin duck, respectively, and Giardia duodenalis assemblage A in a toco toucon. Our results demonstrate that human-pathogenic E. bieneusi genotypes D and Peru6 and G. duodenalis assemblage A are present in captive birds in Brazil, corroborating their potential role as a source of human infection and environmental contamination. [ABSTRACT FROM AUTHOR]

Although cholesterol is essential for membrane fluidity and deformability, the level of its lateral heterogeneity at the plasma membrane of living cells is poorly understood due to lack of appropriate probe. We here report on the usefulness of the D4 fragment of Clostridium perfringens toxin fused to mCherry (theta*), as specific, non-toxic, sensitive and quantitative cholesterol-labeling tool, using erythrocyte flat membrane. By confocal microscopy, theta* labels cholesterol-enriched submicrometric domains in coverslip-spread but also gel-suspended (non-stretched) fresh erythrocytes, suggesting in vivo relevance. Cholesterol domains on spread erythrocytes are stable in time and space, restricted by membrane:spectrin anchorage via 4.1R complexes, and depend on temperature and sphingomyelin, indicating combined regulation by extrinsic membrane:cytoskeleton interaction and by intrinsic lipid packing. Cholesterol domains partially co-localize with BODIPY-sphingomyelin-enriched domains. In conclusion, we show that theta* is a useful vital probe to study cholesterol organization and demonstrate that cholesterol forms submicrometric domains in living cells. [ABSTRACT FROM AUTHOR]

This work aims at developing an active chitosan film through the incorporation of quercetin and the evaluation of physical and functional properties of the films made thereof. The addition of quercetin showed to influence films' properties in terms of surface morphology, tensile strength, and opacity while elongation-at-break, thickness, water vapor, and oxygen permeability were not significantly affected with incorporation of quercetin. The color parameters of chitosan films were affected by quercetin incorporation with a decrease of the values of L* and a*. The film exhibited a high free-radical scavenging activity, showing antioxidant activity. The film-forming solutions of chitosan with or without quercetin showed antibacterial activity against four Gram-negative and three Gram-positive bacteria. These results showed that quercetin incorporation in chitosan-based films has potential to be used as a solution for active food packaging. [ABSTRACT FROM AUTHOR]

This work aims at evaluating the effect of an alginate-chitosan nanomultilayer coating, obtained by electrostatic layer-by-layer self-assembling, in the quality and shelf life of fresh-cut mangoes. Coated and uncoated fresh-cut mangoes were stored under refrigeration (8 °C) for 14 days. The changes in mass loss, titratable acidity, pH, ascorbic acid content, total soluble solids, malondialdehyde content, browning rate, and microbial count were evaluated during storage. At the end of the storage period, lower values of mass loss, pH, malondialdehyde content, browning rate, soluble solids, microorganisms' proliferation, and higher titratable acidity were observed in the coated mangoes. The nanomultilayer coating did not improve the retention of vitamin C during storage of fresh-cut mangoes. Results suggest that chitosan-alginate nanomultilayer edible coating extends the shelf life of fresh-cut mangoes up to 8 days. [ABSTRACT FROM AUTHOR]

This study aimed at the encapsulation of quercetin into lecithin/chitosan nanoparticles using the electrostatic self-assembly technique, followed by evaluation of their functionality (antioxidant activity) and stability at different environmental conditions. These nanoparticles were characterized in terms of: average size, morphology, zeta potential, encapsulation efficiency, loading, and spectroscopic characteristics. Quercetin has been successfully encapsulated in lecithin/chitosan nanoparticles with an efficiency of 96.13 ± 0.44 %. Nanoparticles presented a spherical morphology with an average size of 168.58 ± 20.94 nm and a zeta potential of 56.46 ± 1.94 mV. Stability studies showed that nanoparticles are stable to temperatures ranging between 5 and 70 °C and a pH variation from 3.3 to 5.0. Moreover, encapsulated quercetin showed improved antioxidant properties when compared to free-quercetin. Our results suggest that quercetin-loaded lecithin/chitosan nanoparticles can be used in the manufacture of functional foods. [ABSTRACT FROM AUTHOR]

This work aimed at the characterisation of a nano-laminate coating produced by the layer-by-layer methodology and its evaluation on the preservation of 'Coalho' cheese. Initially, five alternate layers of alginate and lysozyme were assembled in an aminolysed/charged polyethylene terephthalate (A/C PET) and physically characterised by UV/VIS spectroscopy, contact angle, water vapour (WVTR) and oxygen (OTR) transmission rates and scanning electron microscopy. Afterwards, the same methodology was used to apply the nano-laminate coating in 'Coalho' cheese and its shelf life was evaluated during 20 days in terms of mass loss, pH, lipid peroxidation, titratable acidity and microbial count. UV/VIS spectroscopy and contact angle analyses confirmed the layers' deposition and the successful assembly of nano-laminate coating on A/C PET surface. The coating presented WVTR and OTR values of 1.03 × 10 and 1.28 × 10 g m s, respectively. After 20 days, coated cheese showed lower values of mass loss, pH, lipidic peroxidation, microorganisms' proliferation and higher titratable acidity in comparison with uncoated cheese. These results suggest that gas barrier and antibacterial properties of alginate/lysozyme nanocoating can be used to extend the shelf life of 'Coalho' cheese. [ABSTRACT FROM AUTHOR]

Nanolayered coatings of κ-carrageenan, a polysaccharide with good gas barrier properties, and lysozyme, a protein with antimicrobial action, were in a first stage assembled on aminolysed/charged polyethylene terephthalate (PET) pieces, which acted as a support, by alternate five-layer deposition. This was performed to allow the characterization of the nanomultilayer system. PET aminolysis was confirmed by Fourier transform infrared spectroscopy and contact angle, and the subsequent layer adsorption on aminolysed PET surface was confirmed by absorbance, contact angle and SEM images. The water vapour permeability and the oxygen permeability (OP) of the five layers were found to be 0.013 ± 0.003 × 10 and 0.1 ± 0.01 × 10 g m s Pa, respectively. The nanomultilayer system was subsequently applied (without PET support) directly on 'Rocha' ( Pyrus communis L.) fresh-cut pears and whole pears. Uncoated fresh-cut pears and whole pears presented higher mass loss, higher total soluble solids (TSS) and lower titratable acidity when compared with coated fresh-cut pears and whole pears. Uncoated fresh-cut pears also presented a darker colour. These results showed that the nanolayered coating assembled on the fruits' surface has a positive effect on fruit quality and contributed to extend the shelf-life. [ABSTRACT FROM AUTHOR]

Protein deglycation, a new form of protein repair, involves several enzymes. Fructosamine-3-kinase (FN3K), an enzyme found in mammals and birds, phosphorylates fructosamines on the third carbon of their sugar moiety, making them unstable and causing them to detach from proteins. This enzyme acts particularly well on fructose-epsilon-lysine, both in free form and in the accessible regions of proteins. Mice deficient in FN3K accumulate protein-bound fructosamines and free fructoselysine, indicating that the deglycation mechanism initiated by FN3K is operative in vivo. Mammals and birds also have an enzyme designated 'FN3K-related protein' (FN3KRP), which shares ≈65% sequence identity with FN3K. Unlike FN3K, FN3KRP does not phosphorylate fructosamines, but acts on ribulosamines and erythrulosamines. As with FN3K, the third carbon is phosphorylated and this leads to destabilization of the ketoamines. Experiments with intact erythrocytes indicate that FN3KRP is also a protein-repair enzyme. Its physiological substrates are most likely formed from ribose 5-phosphate and erythrose 4-phosphate, which give rise to ketoamine 5- or 4-phosphates. The latter are dephosphorylated by 'low-molecular-weight protein-tyrosine-phosphatase-A' (LMW-PTP-A) before FN3KRP transfers a phosphate on the third carbon. The specificity of FN3K homologues present in plants and bacteria is similar to that of mammalian FN3KRP, suggesting that deglycation of ribulosamines and/or erythrulosamines is an ancient mechanism. Mammalian cells contain also a phosphatase acting on fructosamine 6-phosphates, which result from the reaction of proteins with glucose 6-phosphate. [ABSTRACT FROM AUTHOR]

The aim of this work was to characterise the affinity of protein preparations from Moringa oleifera seeds, specifically extract (seeds homogenised with 0.15 M NaCl), fraction (extract precipitated with 390 mg mL of ammonium sulphate) and cMoL (coagulant M. oleifera lectin) to bind humic acids using a haemagglutinating activity assay with rabbit erythrocytes and a radial diffusion assay in agarose gel. Specific haemagglutinating activity (SHA) decreased by 94 % for the extract and cMoL and by 50 % for the fraction in the presence of humic acid. Precipitation bands were observed in the diffusion gel. Both results suggested humic acid- cMoL binding. Carbohydrates, potassium, and calcium ions and pH affected the SHA of cMoL. As an example of application, cMoL was immobilised on a column packed with sepharose receiving 20 mg mL of carbon humic acid solution, 30 mg of humic acid per gram of support was removed. This result suggested that protein preparations might be used in water treatment to remove humic acids. [ABSTRACT FROM AUTHOR]

2 full factorial design was used to identify the main effects and interactions of the initial medium pH, soybean flour concentration, temperature and orbital agitation speed on extracellular collagenase production by Penicillium aurantiogriseum URM4622. The most significant variables for collagenase production were soybean flour concentration and initial medium pH that had positive main effects, and temperature that had a negative one. Protein concentration in soybean flour revealed to be a significant factor for the production of a collagenase serine proteinase. The most favorable production conditions were found to be 0.75% soybean flour, pH 8.0, 200 rpm, and 28°C, which led to a collagenase activity of 164 U. The enzyme showed an optimum activity at 37°C and pH 9.0, was stable over wide ranges of pH and temperature (6.0 ∼ 10.0 and 25 ∼ 45°C, respectively) and was strongly inhibited by 10 mM phenylmethylsulphonylfluoride. The firstorder rate constants for collagenase inactivation in the crude extract, calculated from semi-log plots of the residual activity versus time, were used in Arrhenius and Eyring plots to estimate the main thermodynamic parameters of thermoinactivation ( E* = 107.4 kJ/mol and ΔH* = 104.7 kJ/mol). The enzyme is probably an extracellular neutral serine collagenase effective on azocoll, gelatin and collagen decomposition. [ABSTRACT FROM AUTHOR]

Background: Morbid obesity is highly prevalent in the Western World, and its consequences present a real public health challenge. Voice alterations can represent one of these consequences and represent an opportunity for interference with therapeutic methods. This particularly features of the individual's voice was the goal of the present study. Methods: A group of 45 adult volunteers of both sexes with a BMI greater than 35 Kg/m was selected among patients of the Obesity Ambulatory of the Digestive Surgery Division. The control group consisted of volunteers matched by sex, age (±1 year), and smoking habits, but with a BMI bellow 30 Kg/m. All subjects were submitted to laryngoscopic examination, audio perceptive analysis, and voice acoustics determination. Examinations were always performed by the same doctor, and diagnoses were provided by two different physician specialists in laryngology and voice. Results: Obese individuals exhibit the following modifications in voice feature: hoarseness, murmuring, vocal instability, altered jitter and shimmer, and reduced maximum phonation times as well the presence of voice strangulation at the end of emission. Conclusion: The voices of individuals with morbid obesity are different of the voice of nonobese people and demonstrate significant changes in vocal characteristics. [ABSTRACT FROM AUTHOR]

Intestine proteases of Nile tilapia ( Oreochromis niloticus) were partially purified by heat treatment (purification factor of 3.5, enzyme activity remained almost constant) to reach the maximum activity and stability within an alkaline pH range of 7.2–11.0. The optimum temperature and stability over a 120 min period were found to be at 55°C and at 35–45°C, respectively. The proteases’ activity was not affected by a 1 vol. % saponin surfactant, inactivated by 0.01 g mL−1 sodium dodecylsulphate after 120 min, and it remained stable for 30 min in a 5 vol. % and 10 vol. % hydrogen peroxide solutions. The proteases were slightly activated by Ca2+, Mg2+, and K+ and the substrate most effectively hydrolysed was casein (40.0 U mg−1). A 24 full factorial design used to evaluated the influence of independent variables showed that the enzyme extract, detergent concentration and the incubation time had a significant influence on the enzymatic activity. The best conditions to be used concerning detergent additive were found with 0.3 mg mL−1 of protein and 3.0 mg mL−1 of detergent for 30 min in the presence of Astrus® detergent. [ABSTRACT FROM AUTHOR]

OBJECTIVE:To assess the effect of bathing with water only or with mild pH neutral soap and water on skin flora of preterm newborns.METHOD:Randomized blinded clinical trial that enrolled 73 preterm newborns whose gestational age was between 28 and 35 weeks and birth weight between 800 and 1800?g. At the age of 3 days the infants were randomly assigned to a group that was given daily baths either with water only, or with soap and water for 7 or more days. Antibiotic treatment delayed onset of the trial in some preterm infants. On the final day of bathing axillary bacterial cultures were obtained before and 30 minutes after bathing.RESULTS:At the time of axillary culturing, the mean age of the newborns was 19 days. Coagulase-negative Staphylococcus was the most prevalent microorganism in both groups. No significant difference was found in the count of microorganism colonies between both groups. The comparison of the number of bacterial colonies between the time before and after bath, carried out by repeated-measures ANOVA showed a significant difference over time in the two groups, without a significant difference between the two groups.CONCLUSIONS:Bathing preterm newborns with water only or with soap and water produces similar effects on skin colonization of a preterm neonate. Both are effective to decrease the number of colonies of Gram-positive and Gram-negative bacteria.Journal of Perinatology (2005) 25, 375-379. doi:10.1038/sj.jp.7211332 Published online 28 April 2005 [ABSTRACT FROM AUTHOR]

We report the identification of the mutations in the only known case of L-3-phosphoserine phosphatase deficiency, a recessively inherited condition. The two mutations correspond to the replacement of the semiconserved Asp32 residue by an asparagine and of the extremely conserved Met52 by a threonine. The effects of both mutations were studied on the human recombinant enzyme, expressed in Escherichia coli. Met52Thr almost abolished the enzymatic activity, whereas the Asp32Asn mutation caused a 50% decrease in Vmax. In addition, L-serine, which inhibits the conversion of [14C] phosphoserine to serine when catalysed by the wild-type enzyme, had a lesser inhibitory effect on the Asp32Asn mutant, indicating a reduction in the rate of phosphoenzyme hydrolysis. These modifications in the properties of the enzyme are consistent with the modification in the kinetic properties observed in fibroblasts from the patient.European Journal of Human Genetics (2004) 12, 163-166. doi:10.1038/sj.ejhg.5201083 Published online 3 December 2003 [ABSTRACT FROM AUTHOR]

Unlabelled: Glucose-6-phosphatase is a multicomponent enzymatic system of the endoplasmic reticulum, which catalyses the terminal steps of gluconeogenesis and glycogenolysis by converting glucose-6-phosphate to glucose and inorganic phosphate. Glycogen storage diseases type I (GSD I) are a group of metabolic disorders arising from a defect in a component of this enzymatic system, i.e. the glucose-6-phosphate hydrolase (GSD Ia), the glucose-6-phosphate translocase (GSD Ib) and possibly also the translocases for inorganic phosphate (GSD Ic) or glucose (GSD Id). The genes encoding the glucose-6-phosphate hydrolase and the glucose-6-phosphate translocase have both been cloned and assigned to human chromosomes 17q21 and 11q23, respectively. Investigation of patients with GSD I shows that those with GSD Ia are mutated in the glucose-6-phosphate hydrolase gene, whereas those diagnosed as GSD Ib, GSD Ic or GSD Id are mutated in the glucose-6-phosphate translocase gene, and are therefore GSD Ib patients, in agreement with the fact that they all have neutropenia or neutrophil dysfunction. This suggests that the biochemical assays used to differentiate GSD Ic and GSD Id from GSD Ib are not reliable.Conclusion: In practice therefore appears to be only two types of GSD I (Ia and Ib), which can be differentiated by (1) measurement of glucose-6-phosphatase activity in fresh and detergent-treated homogenates and (2) by mutation search in the genes encoding the glucose-6-phosphate hydrolase and the glucose-6-phosphate translocase. [ABSTRACT FROM AUTHOR]

The purpose of this work was to test the hypothesis that mutations in the putative glucose 6-phosphate translocase gene would account for most of the cases of GSD I that are not explained by mutations in the phosphohydrolase gene, ie that are not type Ia. Twenty-three additional families diagnosed as having GSDI non-a (GSD Ib, Ic or Id) have now been analysed. The 9 exons of the gene were amplified by PCR and mutations searched both by SSCP and heteroduplex analysis. Except for one family in which only one mutation was found, all patients had two allelic mutations in the gene encoding the putative glucose 6-phosphate translocase. Sixteen of the mutations are new and they are all predicted to lead to nonfunctional proteins. All investigated patients had some degree of neutropenia or neutrophil dysfunction and the clinical phenotype of the four new patients who had been diagnosed as GSD Ic and the one diagnosed as GSD Id was no different from the GSD Ib patients. Since these patients, and the four type Ic patients from two families previously studied, shared several mutations with GSD Ib patients, we conclude that their basic defect is in the putative glucose 6-phosphate translocase and that they should be reclassified as GSD Ib. Isolated defects in microsomal Pi transporter or in microsomal glucose transporter must be very rare or have phenotypes that are not recognised as GSD I, so that in practice there are only two subtypes of GSD I (GSD Ia and GSD Ib). [ABSTRACT FROM AUTHOR]

In the original publication, article title was incorrectly published. [ABSTRACT FROM AUTHOR]