Search

As with other pathogens, competitive interactions between Bordetella pertussis strains drive infection risk. Vaccines are thought to perturb strain diversity through shifts in immune pressures; however, this has rarely been measured because of inadequate data and analytical tools. We used 3344 sequences from 23 countries to show that, on average, there are 28.1 transmission chains circulating within a subnational region, with the number of chains strongly associated with host population size. It took 5 to 10 years for B. pertussis to be homogeneously distributed throughout Europe, with the same time frame required for the United States. Increased fitness of pertactin-deficient strains after implementation of acellular vaccines, but reduced fitness otherwise, can explain long-term genotype dynamics. These findings highlight the role of vaccine policy in shifting local diversity of a pathogen that is responsible for 160,000 deaths annually.

Vegetables are potential vehicles for the transmission of parasites. Helminth eggs have been detected in produce in various countries around the world, with high prevalence rates. However, there is no standard validated method or ISO to the isolation, concentration, and identification of helminth eggs in salad vegetables, which makes comparison difficult since each study utilized different analytical methods and food matrix. The purpose of the present study was to evaluate the effectiveness of a previously described methodology developed for the recovery of Ascaris sp. eggs in two different leaf greens under real conditions of use, through the simultaneous application of the same method by ten different laboratories. This is the first collaborative inter-laboratory trial by several laboratories across Brazil. Lettuce and arugula samples artificially contaminated with A. suum eggs were analyzed by various laboratories to determine the recovery efficiency of Ascaris sp. eggs. A final mean recovery efficiency of 57.10% for Ascaris sp. eggs in lettuce and 50.7% in arugula was identified. The methodology applied in this study can be used to evaluate Ascaris sp. eggs in vegetables. It is reasonably easy, fast, and efficient in comparison with other methods. The identification of helminth eggs in fresh vegetables using a reliable methodology is important if the microbiological safety of these foods is to be improved.

As with other pathogens, competitive interactions between Bordetella pertussis strains drive infection risk. Vaccines are thought to perturb strain diversity through shifts in immune pressures; however, this has rarely been measured because of inadequate data and analytical tools. We used 3344 sequences from 23 countries to show that, on average, there are 28.1 transmission chains circulating within a subnational region, with the number of chains strongly associated with host population size. It took 5 to 10 years for B. pertussis to be homogeneously distributed throughout Europe, with the same time frame required for the United States. Increased fitness of pertactin-deficient strains after implementation of acellular vaccines, but reduced fitness otherwise, can explain long-term genotype dynamics. These findings highlight the role of vaccine policy in shifting local diversity of a pathogen that is responsible for 160,000 deaths annually.

BackgroundWorldwide, healthcare-associated SARS-CoV-2 infections are a major problem: they are associated with increased morbidity, mortality, and hospitalization costs. In-depth studies across the pandemic are crucial to understand and prevent transmission in hospital settings. The principal aims of this study were to characterise patients and validate ECDC definitions of healthcare-associated COVID-19 infections.MethodsWe set up a retrospective observational study spanning the first three waves of the COVID-19 pandemic in a Belgian university hospital: it describes the characteristics of COVID-19 patients admitted, with either healthcare- or community-associated infections. We performed a cluster analysis through epidemiological and viral genome analyses of the healthcare-associated infections, in order to validate the ECDC definitions of healthcare-associated COVID-19 infections.ResultsBetween week 10 of 2020 and week 22 of 2021, 168 patients were hospitalized with healthcare-associated COVID-19. The following factors were found more often in symptomatic healthcare- than in community-associated hospitalized patients: older age, increased frailty, smoking habits, and comorbidities. The genome-based cluster analyses showed that different viral lineages predominated in different timeframes. We observed a good correlation of epidemiological data with genome sequencing results in at least 12 different outbreaks in our hospital, thus validating the ECDC definitions. ConclusionsThis in-depth characterization sheds new light on the problem of healthcare-associated COVID-19 infections, in particular on patients’ characteristics, epidemiology, and cluster dynamics. Even though epidemiological evaluation of nosocomial infections is vital, management of nosocomial outbreaks can undoubtedly benefit from genome sequencing analyses to reinforce their strategy.

Competitive interactions between pathogen strains drive infection risk. Vaccines are thought to perturb strain diversity through shifts in immune pressures, however, this has rarely been measured due to inadequate data and analytical tools. Bordetella pertussis (B. pertussis), responsible for 160,000 deaths annually1, provides a rare natural experiment as many countries have switched from whole cell vaccines to acellular vaccines, which have very different immunogenic properties2,3. Here we use 3,344 sequences from 23 countries and build phylogenetic models to reveal that B. pertussis has substantial diversity within communities, with the relative fitness of local genotypes changing in response to switches in vaccine policy. We demonstrate that the number of transmission chains circulating within subnational regions is strongly associated with host population size. It takes 5-10 years for individual lineages to be homogeneously distributed throughout Europe or the United States. Increased fitness of pertactin-deficient strains following implementation of acellular vaccines, but reduced fitness otherwise, can explain long-term genotype dynamics. These findings highlight the role of national vaccine policies in shifting local diversity of a pathogen that still poses a large burden on global public health.

In Western Europe, the incidence of both respiratory and cutaneous diphtheria, caused by toxin-producing Corynebacterium diphtheriae, Corynebacterium ulcerans or Corynebacterium pseudotuberculosis, has been low over the past few decades thanks to the use of an effective vaccine and a high level of vaccination coverage. However, the disease has still not been eradicated and continues to occur in all of Europe. In order to prevent sequelae or a fatal outcome, diphtheria antitoxin (DAT) should be administered to suspected diphtheria patients as soon as possible, but economic factors and issues concerning regulations have led to poor availability of DAT in many countries. The European Centre for Disease Prevention and Control and World Health Organization have called for European Union-wide solutions to this DAT-shortage. In order to illustrate the importance of these efforts and underline the need for continued diphtheria surveillance, we present data on all registered cases of toxigenic and non-toxigenic C. diphtheriae, C. ulcerans and C. pseudotuberculosis in Belgium during the past decade, up to and including 2017.

BackgroundAn outbreak of diphtheria, declared in Yemen in October 2017, is still ongoing. Methods. Probable cases were recorded through an electronic diseases early warning system. Microbiological culture, genomic sequencing, antimicrobial susceptibility and toxin production testing were performed.MethodsProbable cases were recorded through an electronic diseases early warning system. Microbiological culture, genomic sequencing, antimicrobial susceptibility and toxin production testing were performed.FindingsThe Yemen diphtheria outbreak developed in three epidemic waves, which affected nearly all governorates (provinces) of Yemen, with 5701 probable cases and 330 deaths (October 2017 - April 2020). The median age of patients was 12 years (range, 0.17-80). Virtually all outbreak isolates (40 of 43 tested ones) produced the diphtheria toxin. We observed low level of antimicrobial resistance to penicillin. We identified six separate Corynebacterium diphtheriae phylogenetic sublineages, three of which are genetically related to isolates from Saudi Arabia and Somalia. The predominant sublineage was resistant to trimethoprim and was associated with unique genomic features, more frequent neck swelling (p=0.002) and a younger age of patients (p=0.06). Its evolutionary rate was estimated at 1.67 × 10−6 substitutions per site year-1, placing its most recent common ancestor in 2015, and indicating silent circulation of C. diphtheriae in Yemen earlier than outbreak declaration.InterpretationWe disclose clinical, epidemiological and microbiological characteristics of one of the largest contemporary diphtheria outbreaks and demonstrate clinically relevant heterogeneity of C. diphtheriae isolates, underlining the need for laboratory capacity and real-time microbiological analyses to inform prevention, treatment and control of diphtheria.FundingThis work was supported by institutional funding from the National Centre of the Public Health Laboratories (Sanaa, Yemen) and Institut Pasteur (Paris, France) and by the French Government Investissement d ‘Avenir Program.

Improving the aerodynamics of heavy trucks is an important consideration in the strive for more energy-efficient vehicles. Cooling drag is one part of the total aerodynamic resistance acting on a vehicle, which arises as a consequence of air flowing through the grille area, the heat exchangers, and the irregular under-hood area. Today cooling packages of heavy trucks are dimensioned for a critical cooling case, typically when the vehicle is driving fully laden, at low speed up a steep hill. However, for long-haul trucks, mostly operating at highway speeds on mostly level roads, it may not be necessary to have all the cooling airflow from an open-grille configuration. It can therefore be desirable for fuel consumption purposes, to shut off the entire cooling airflow, or a portion of it, under certain driving conditions dictated by the cooling demands. In Europe, most trucks operating on the roads are of cab-over-engine type, as a consequence of the length legislations present. However, there are new directions from the European Union, which would permit slightly longer vehicles to improve aerodynamics and also to allow for a safer, more environmentally friendly vehicle. The truck design, where a cab-over-engine cab has an elongated front, is often referred to as a Soft Nose, where, as the name implies, the nose should be “soft” to improve the safety for pedestrians and also for car occupants in the event of a collision. This paper deals with the analysis of cooling airflow for two different front-end designs of a heavy truck. The first design is a cab-over-engine cab; the second is a Soft Nose cab, which in this case is basically an elongation of the grille area of the cab-over-engine cab to obtain a smoother shape of the cab. The Soft Nose model used in this investigation was extended 200 mm from the cab-over-engine front. Computational Fluid Dynamics was used as the tool for examining the aerodynamic properties of the vehicle models. A steady RANS-based approach was conducted, based on the method evaluated in previous work performed by the authors. The cab-over-engine and Soft Nose models were evaluated in an open-road environment. The configurations were evaluated both with inactive and active heat exchangers, in order to examine the effect of heating the air on the drag co-efficient and also to determine the cooling capacity of the different models. A sub- study was performed where different opening percentages of the grille area was investigated to determine the minimum percentage opening that would be needed to achieve a radiator Top Tank Temperature value below a target limit of 100 °C. The results show that there was potential for drag reductions for the Soft Nose model used. The cooling airflow was different for the cab-over-engine and Soft Nose models; as a consequence of the longer distance between the grille and cooling package, less air entered the cooling module for the Soft Nose model. A large portion of the airflow entering the grille leaked around the cooling module for the Soft Nose model. Also, following on from the reduced airflow through the cooling package, the radiator Top Tank Temperature values were considerably increased with the Soft Nose model. It was also shown that for the specific driving condition simulated here, an opening of 17.5% of the grille area was required to ensure sufficient cooling capacity. An interesting continuation of the cooling airflow analysis of the Soft Nose model would be to add ducts, guiding the air from the grille to the cooling module, to investigate if leakage could be reduced and cooling capacity increased.

Objectives Quantifying IgG antibodies to pertussis toxin (PT) is the most specific and sensitive method for the serodiagnosis of a Bordetella pertussis infection. Since PT is a component of acellular pertussis vaccines, anti-PT IgG is also induced by vaccination, precluding pertussis serodiagnosis based exclusively on anti-PT IgG in recently vaccinated subjects. Here, we aim to identify additional B. pertussis-specific serological markers that can discriminate between infection and recent vaccination. Methods The clinical usefulness of measuring IgA directed to the vaccine antigen PT and IgG directed to non-vaccine antigens (Fim2/3, LPS, ACT, CatACT) was evaluated in nine well characterized subject groups, aged 10–89 years (n = 390). Serum anti-PT IgG levels (>125 IU/mL) served as an indicator for a recent B. pertussis infection. Comparing symptomatic pertussis-infected subjects (n = 140) with recently vaccinated, non-infected subjects (n = 100) revealed the optimal cut-off, accuracy, sensitivity and specificity for each single parameter. Results For pertussis diagnosis in recently vaccinated subjects, the measurement of anti-PT IgA (cut-off 15 IU/mL) and anti-ACT IgG (cut-off 15 U/mL) resulted in accuracies of 95% (91.5–97.1) and 87.5% (82.7–91.1), sensitivities of 92.9% (87.4–96.0) and 83.6% (76.5–88.8) and specificities of 98% (93.0–99.4) and 93% (86.3–96.6), respectively. Comparing anti-PT IgA levels between the youngest (10–19 years, n = 38) and oldest (70–89 years, n = 17) age groups revealed an age-dependent increase in antibody levels in pertussis-infected subjects (p Conclusions Reflex testing of anti-PT IgA and anti-ACT IgG improves pertussis serodiagnosis in recently vaccinated symptomatic subjects with elevated anti-PT IgG levels. Furthermore, both markers can discriminate between vaccination and recent infection in pertussis serosurveillance studies.

SUMMARYIn 2015, the Belgian National Reference Centre for Bordetella analyzed 4110 respiratory samples by qPCR and 4877 serum samples by serology. Whereas about 50% of respiratory samples were from infants and children below the age of five, serum samples were distributed among all age categories. A total of 394 (9·6%) cases was diagnosed as positive for Bordetella pertussis by qPCR and 844 (17·3%) cases were diagnosed as acute infection by serology (anti-pertussis toxin (PT) IgG > 125 IU/ml). Another 1042 (21·4%) sera had anti-PT IgG between 55 and 125 IU/ml reflecting a vaccination or pertussis infection during the last 1–2 years. Seventy per cent of the pertussis cases diagnosed by qPRC were in infants and children younger than 14 years old, whereas the highest number of sera with anti-PT levels >125 IU/ml was in the age group of 10–14 years old. Based on the limited data of the last vaccination (reported for only 15% of the samples), recent booster vaccination in the teenager group may have contributed only minimally to these elevated anti-PT levels. The highest number of sera with anti-PT titers between 55 and 125 IU/ml was found in the age category 50–59 years old. It is clear that pertussis continues to be a problem in Belgium and that other vaccination strategies (maternal vaccination, cocoon vaccination) and ultimately better vaccines will be needed to control this highly infectious respiratory disease.

Nowadays, much focus for vehicle manufacturers is directed towards improving the energy efficiency of their products. The aerodynamic drag constitutes one major part of the total driving resistance for a vehicle travelling at higher speeds. In fact, above approximately 80km/h the aerodynamic drag is the dominating resistance acting on a truck. Hence the importance of reducing this resistance is apparent. Cooling drag is one part of the total aerodynamic drag, which arises from air flowing through the heat exchangers, and the irregular under-hood area.When using Computational Fluid Dynamics (CFD) in the development process it is of great importance to ensure that the methods used are accurately capturing the physics of the flow. This paper deals with comparative studies between CFD and wind-tunnel tests.In this paper, two comparative studies are presented. One is a comparison between cooling performance simulations and chassis dynamometer measurements; the other study is a comparison between external aerodynamics simulations and wind-tunnel measurements.The purpose of this study was to evaluate and develop methods and models for determining aerodynamic drag and cooling performance, which ultimately will be used to develop more energy-efficient cooling of heavy trucks.The results from the two comparative studies showed that there was in general good agreement between CFD and wind-tunnel measurements. For the cooling performance simulations, the analysed parameters were very close to the measured values. For the external aerodynamics simulations, the results were not easy to analyse. The overall results were still satisfactory; for the simulated yaw angles, the drag coefficient (CD) values were less than 4.1% different from measured data.

Natural products, including those derived from plants, have largely contributed to the development of therapeutic drugs. Glutamate is the main excitatory neurotransmitter in the central nervous system and it is also considered a nociceptive neurotransmitter, by acting on peripheral nervous system. For this reason, in this study we investigated the effects of the hydroalcoholic extracts from Drymis winteri (polygodial and drimanial), Phyllanthus (rutin and quercetine), Jathopha elliptica (jatrophone), Hedyosmum brasiliense (13HDS), Ocotea suaveolens (Tormentic acid), Protium kleinii (alphabeta-amyrin), Citrus paradise (naringin), soybean (genistein) and Crataeva nurvala (lupeol), described as having antinociceptive effects, on glutamatergic transmission parameters, such as [(3)H]glutamate binding, [(3)H]glutamate uptake by synaptic vesicles and astrocyte cultures, and synaptosomal [(3)H]glutamate release. All the glutamatergic parameters were affected by one or more of these compounds. Specifically, drimanial and polygodial presented more broad and profound effects, requiring more investigation on their mechanisms. The putative central side effects of these compounds, via the glutamatergic system, are discussed.

Extracellular guanine-based purines, namely the nucleotides GTP, GDP, GMP and the nucleoside guanosine, exert important neuroprotective and neuromodulator roles in the central nervous system, which may be related to inhibition of the glutamatergic neurotransmission activity. In this study, we investigated GMP effects on mice inhibitory avoidance performance and the dependence on its conversion to guanosine for such effect, by using the ecto-5'-nucleotidase specific inhibitor AOPCP. We also investigated if this conversion occurs in the central nervous system or peripherally, and if guanosine and GMP affect nociception by the tail-flick test. I.p. GMP or guanosine (7.5 mg/kg) or i.c.v. GMP (480 nmol) pretraining administration was amnesic for the inhibitory avoidance task. I.c.v. AOPCP (1 nmol) administration completely reversed the amnesic effect of i.c.v. GMP, but not of i.p. GMP, indicating that peripheral conversion of GMP to guanosine is probably relevant to this effect. AOPCP alone did not interfere with the performance. Furthermore, tail-flick measurement was unaffected by i.p. GMP and guanosine, suggesting that the amnesic effect of both purines was not due to some antinociceptive effect against the footshock used in the task. All these data together, in accordance to those previously observed in studies involving glutamate uptake and seizures reinforce the idea that guanosine is the specific extracellular guanine-based purines effector and indicate that its conversion occurs not only in the central nervous system but also peripherally.

Glutamate is to be considered a nociceptive neurotransmitter and glutamatergic antagonists present antinoceptive activity. In this study we investigated the effects of the naturally occurring antinociceptive compounds rutin, geraniin and quercetine extracted from Phyllanthus, as well as the diterpene jatrophone, extracted from Jatropha elliptica on the binding of [3H]glutamate and [3H]GMP-PNP [a GTP analogue which binds to extracellular site(s), modulating the glutamatergic transmission] in rat brain membrane. Jatrophone inhibited [3H]glutamate binding and geraniin inhibited [3H]GMP-PNP binding. Quercetine inhibited the binding of both ligands. These results may indicate a neurochemical parameter possibly related to the antinoceptive activity of these natural compounds.

Metabotropic glutamate receptors (mGluRs) have been shown to modulate adenylate cyclase activity via G-proteins. In the present study we report similar results to the previously observed in the literature, showing that glutamate and the metabotropic agonists, 1S,3R-ACPD or quisqualate induced cAMP accumulation in hippocampal slices of young rats. Moreover, guanine nucleotides GTP, GDP or GMP, inhibited the glutamate-induced cAMP accumulation. By measuring LDH activity in the buffer surrounding the slices, we showed that the integrity of the slices was maintained, indicating that the effect of guanine nucleotides was extracellular. GMP, GDPβ-S or Gpp(NH)p abolished quisqualate-induced cAMP accumulation. GDPβ-S or Gpp(NH)p but not GMP inhibited 1S,3R-ACPD-induced cAMP accumulation. The response evoked by glutamate was also abolished by the mGluR antagonists: L-AP3 abolished glutamate-induced cAMP accumulation in a dose-dependent manner and MCPG was effective only at the 2 mM dose. DNQX was ineffective. We are reporting here, an inhibition induced by guanine nucleotides, via an extracellular site (s), similar to the observed with classical glutamate antagonists on a cellular response evoked by mGluR agonists.

Today there are a large variety of drag reducing devices for heavy trucks that are commonly used, for example roof deflectors, cab side extenders and chassis fairings. These devices are often proven to be efficient, reducing the total aerodynamic resistance for the vehicle. However, the drag reducing devices are usually identical for a specific pulling vehicle, independent of the layout of the vehicle combination. In this study, three vehicle combinations were analyzed. The total length of the vehicles varied between 10.10m and 25.25m. The combinations consisted of a rigid truck in combination with one or two cargo units. The size of the gap between the cargo units differed between the vehicle combinations. There were also three configurations of each vehicle combination with different combinations of roof deflector and cab side extenders, yielding a total number of nine configurations. The aim of this investigation was to determine the aerodynamic effects of the roof deflector and cab side extenders as a function of the type of vehicle combination. Important factors were the total length of the vehicle combination and the influence of the drag reducing devices further downstream. The investigation was performed using Computational Fluid Dynamics (CFD). The results from the investigation showed that the effect of the two drag reducing devices analyzed was different depending on the type of vehicle combination. It was established that the roof deflector and cab side extenders were always efficient in reducing drag both in 0° yaw and 5° yaw but the magnitudes differed between the configurations. The largest effects of the drag reducing devices were seen for the truck including the 1st cargo unit; the influence of the drag reducing devices on the 2nd cargo unit was smaller. The aerodynamic reductions were diminishing downstream, why it was concluded that it is of great importance to improve the aerodynamic design of the rest of the vehicle to maintain the positive effects of the drag reducing devices.

The amount of cotyledon polyphenolic cells varies extensively within the Theobroma species. The polyphenolic compounds of these cells play a protective role and furthermore have an important function in the development of chocolate flavour. The morphology of the polyphenolic cells of the mesophyll is described and the development of these idioblasts in Theobroma cacao L., T. subincanum Mart., T. obovatum Klotzsch ex Bernoulli, T. grandiflorum (Willd. ex Spreng.) K. Schum., T. microcarpum Mart., T. bicolor Bonpl. and T. speciosum Willd. ex Spreng analysed. The total polyphenolic content in the seeds as determined by spectrophotometry showed a variation of about forty times. The alive, transparent polyphenolic cells are scattered throughout the cotiledonary mesophyll. However the polyphenolic cells of T. cacao and T. grandiflorum are also aligned perpendicularly with respect to the mesophyll borders and, in addition, both species display polyphenolic cells with a natural translucent purple colour. All the species analysed contained polyphenolic cells scattered throughout the parenchymal cells and also in a lengthwise association with vascular bundles. In T. bicolor and T. speciosum, the species with the lowest polyphenolic contents, these cells were mostly located around the vascular bundles. Using Scanning Electron Microscopy, the polyphenolic cells demonstrated a complex cytoarchitecture, and after fixing with glutaraldhyde, the polyphenolic secretion was shown to remain as a single unit or was organized into round droplets. Transmission Electron Microscopy displayed immature plastids from young mesophyll cells containing eletron-dense deposits similar to phenolic substances, suggesting that Theobroma plastids are involved in the synthesis of phenolics. A quantidade de células polifenólicas dos cotilédones das espécies de Theobroma variam extensivamente. Os compostos polifenólicos destas células além da função de proteção participam no desenvolvimento do aroma do chocolate. A morfologia das células polifenólicas do mesofilo é descrita e também é analisado o desenvolvimento dos idioblastos em T. cacao L., T. subincanum Mart., T. obovatum Klotzsch ex Bernoulli, T. grandiflorum (Willd. ex Spreng.) K. Schum., T. microcarpum Mart., T. bicolor Bonpl. e T. speciosum Willd. ex Spreng. A quantidade de polifenóis totais determinada por espectrofotometria apresentou uma grande variação, cerca de 40 vezes entre T. cacao (variedade forastero). As células polifenólicas apresentam-se transparentes e dispersas pelo mesofilo cotiledonar. Em T. cacao e T. grandiflorum grande número de células estão alinhadas perpendicularmente à borda do mesofilo e apresentam coloração púrpura translúcida. Entretanto, em todas as espécies as células polifenólicas também associam-se ao feixe vascular. Nas espécies T. bicolor e T. speciosum, com baixo conteúdo polifenólico, estão localizadas apenas na região do feixe vascular. À microscopia eletrônica de varredura, as células polifenólicas apresentam uma complexa citoarquitetura e após a fixação em glutaraldeído, a secreção polifenólica exibe um só conteúdo ou fragmenta-se em várias gotas esféricas. À microscopia eletrônica de transmissão aparecem nas células jovens do mesofilo, plastídeos imaturos eletrondensos, semelhantes a substâncias polifenólicas sugerindo que em Theobroma os plastídeos estão envolvidos na síntese de polifenóis.

Localization of the cotyledon reserves of Theobroma grandiflorum (Willd. ex Spreng.) K. Schum., T. subincanum Mart., T. bicolor Bonpl. and their analogies with T. cacao L.). Cotyledon mesophyll cell morphology and lipid and protein synthesis of T. grandiflorum, T. subincanum and T. bicolor were analyzed and compared with T. cacao. These species possess foliar cotyledons folded around the hypocotyl radicle axis, typical of Sterculiaceae. Fruit size, morphology and weight are very distinct amongst the four species and so are the respective seeds. The main axis of the T. grandiflorum and T. bicolor seeds measured about 30 mm, while T. subincanum and T. cacao seeds measured 17 mm and 26 mm respectively. The seed weights of T. grandiflorum, T. bicolor, T. subincanum and T. cacao were 11.6 g, 9.4 g, 2.1 g and 3.0 g, respectively. The cotyledon mesophylls of the four species contained mainly polysaccharides and lipid-protein reserve cells. Theobroma cacao, T. grandiflorum and T. subincanum were composed of greater than 50% lipids. For the four species, lipid globules gradually accumulated adjacent to the cell wall, and these globules measured from 1 to 3 µm. TEM showed low-density proteins inside the central vacuole of the young mesophyll cells of T. cacao. The protein reserves of the mature cells were densely scattered amongst the lipid bodies, and a few starch granules occurred together with the cotyledon mesophyll of the four species. Polyphenolic cells were found throughout the mesophyll cells or aligned with the respective vascular bundles. Immature cells demonstrated the capacity to synthesize all these reserves, but gradually the pre-determined cells produced mainly lipid-protein reserves. Besides the unique characteristics of the T. cacao products, the lipid-protein synthesis capacities of T. grandiflorum, T. subincanum and T. bicolor suggest various possibilities for new industrialized food, pharmaceutical and cosmetic products.

Natural products including those derived from plants, have over the years greatly contributed to the development of therapeutic drugs. Polygodial and drimanial are sesquiterpenes isolated from the bark of the plant Drymis Winteri (Winteraceae) that exhibit antinociceptive properties. Since peripheral glutamate presents nociceptive actions, in this study it was investigated the effects of hydroalcooholic extracts from Drymis winteri (polygodial and drimanial) on the glutamatergic system in rat brain. Polygodial and drimanial inhibited glutamate uptake by astrocytes, as well as by cortical, hippocampal and striatal slices, and increased synaptosomal glutamate release. These concurrent effects would predispose to an increase in the extracellular glutamate concentrations, leading to possible neurotoxic effects (excitotoxicity) of these natural compounds, which would suggest the need for some caution in their therapeutic application.

Glutamatergic mechanisms are thought to be involved in stress-induced changes of brain function, especially in the hippocampus. We hypothesized that alterations caused by the hormonal changes associated with chronic and acute stress may affect glutamate uptake and release from hippocampal synaptosomes in Wistar rats. It was found that [3H]glutamate uptake and release by hippocampal nerve endings, when measured 24 h after 1 h of acute restraint, presented no significant difference. The exposure to repeated restraint stress for 40 days increased neuronal presynaptic [3H]glutamate uptake as well as basal and K+-stimulated glutamate release when measured 24 h after the last stress session. Chronic treatment also caused a significant decrease in [3H]glutamate binding to hippocampal membranes. We suggest that changes in the glutamatergic system are likely to take part in the mechanisms involved in nervous system plasticity following repeated stress exposure.

Besides their well-defined intracellular roles in transmembrane signals transduction, guanine derivatives play important roles by acting from the outside of neural cell membranes. These roles are mediated by two different pool sites in cell membranes: G proteins, which bind to specific (GDP and GTP) intracellular guanine derivatives, and sites that bind to extracellular guanine derivatives. In this study we investigated some methodological characteristics of both guanine derivatives binding sites (intracellular and extracellular) in rat brain neural membranes. By investigating the binding of a poorly hydrolyzed GTP analogue and the adenylate cyclase activity in neural membranes, we observed some distinctiveness of guanine derivatives binding sites: stability to washing procedures (extracellular) and modulation of adenylate cyclase activity (intracellular). These results allow dealing with each site separately, which could be useful for discriminating the roles of extracellular and intracellular guanine derivatives in the central nervous system.

Orientador : Debora de Queiroz Tavares Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos Resumo: Nos estudos sobre as reservas do mesofilo cotiledonar do gênero Theobroma destacam-se os trabalhos sobre sementes de T. cacao, fonte única da matéria prima do chocolate. Estas pesquisas preferenciam as mudanças ocorridas na estrutura celular durante a fermentação, secagem, torração; etapas do processo de produção do chocolate. Mais recentes são os trabalhos com sementes de T. grandiflorum utilizadas na elaboração do cupulate, produto com características semelhantes ao chocolate. O presente trabalho analisa as células de reserva lípide-proteicas e idioblastos de polifenóis de T. cacao, T. grandiflorum, T. subincanum, T. bicolor e T. speciosum. Em T. obovatum e T. microcarpum o estudo apresentou apenas as células polifenólicas. A distribuição dos glóbulos lipídicos nas células são análogas nestas espécies, isto é, localização periférica e extrema susceptibilidade ao calor, coalescendo sob condições térmicas acima de 30°C. A proteína de reserva apresenta-se entremeada com os glóbulos lipídicos e alguns grãos de amido. As células polifenólicas, em T. bicolor e T. speciosum, localizam-se na região do feixe vascular e parênquima da radícula e nas outras espécies estudadas estão dispersas pelo mesofilo e próximas ao feixe vascular. A mucilagem é uma secreção celular constante em cinco espécies estudadas, não se acumulando dentro das células, porém com raras exceções ocorrem células com mucilagem na epiderme do mesofilo (T. subincanum). T. speciosum caracteriza-se por efetivamente apresentar células de mucilagem. Estas células são abundantes no mesofilo e cerca de 10% alcançam grandes dimensões sem rompimento do tecido. As células polifenólicas apresentam características histoquímicas semelhantes a mucilagem, porém sua estreita relação com os feixes vasculares direciona seu desenvolvimento para a síntese de polifenóis. O tecido do mesofilo cotiledonar é semelhante em todas as espécies aqui estudadas devido a predominância de reserva lípide-proteica. Células imaturas de sementes verdes demonstram capacidade para sintetizar todas as reservas; na maturidade celular demonstram predominância de reservas lípide-proteicas ou de mucilagem ou de polifenóis. A distribuição das reservas no mesofilo aproximam T. grandiflorum à T. subincanum. A maior quantidade de proteínas está em T. bicolor. Esta espécie apresenta menor quantidade de lipídeos e de células polifenólicas Abstract: Many investigations on the cellular reserves within the cotyledon mesophyll of the Theobroma have been carried out on Theobroma cacao seeds, the source of raw for the production of chocolate. These studies demonstrated structural cellular changes due to fermentation, drying, and roasting processes during chocolate production. Recently the seeds of the T. grandiflorum have been analyzed due to their importance in the production of ¿cupulate¿, a new product similar to chocolate. In the present work, we analyze the lipid-protein reserve cells and polyphenolic idioblasts derived from T. cacao, T. grandiflorum, T. subincanum, T. bicolor, T. speciosum and T. obovatum. Only phenolic cells were studied in T.obovatum and T. microcarpum.The distribution of lipid bodies was similar in all the species studied; that is, they were located at the periphery of the cell wall and demonstrated extremely susceptibly to heat, coalescing at temperatures above 30°C. The reserve protein was interspersed with lipid bodies and some starch granules. The polyphenolic cells were dispersed throughout the mesophyll and around the vascular bundles in all the species studied. In T. bicolor and T. speciosum the polyphenolic cells were only observed around the vascular bundle and radicle parenchyma. Mucilage is a cellular secretion in the five species studied. Mucilage does not accumulate intracellularly; however, rarely we did observe mucilage in the cells of the mesophyll epidermis of T. subincanum. In T. speciosum, mucilage cells were abundant in the mesophyll and about 10% reached large dimensions without tissue disruption. The polyphenolic cells presented characteristic mucilage histochemistry, however their strict association with vascular bundles should direct their development towards polyphenol synthesis. Cotyledon mesophyll tissue is similar in all the species studied with respect to lipid-protein reserve. Immature cells demonstrated the capacity to synthesize all the reserves, but this capacity is not retained in the mature cell. The distribution of reserves in the mesophyll are most similar in T. grandiflorum and T. subincanum. T. bicolor showed a high protein content, lower lipid levels and low levels of polyphenolic cells Doutorado Doutor em Alimentos e Nutrição

We examined samples of irrigation water and vegetables from kitchen gardens in Campinas, Brazil. The bacterial analysis condemned 22.3% of the vegetable samples, and the parasitological examination condemned 14.5%. The criteria established by the Brazilian legislation condemned 11.8% of the irrigation water samples. Parasites were significantly more frequent in vegetables in the rainy season, while excessive fecal coliforms were more frequent in the dry season. A proper monitoring of the irrigation water supply is important to avoid the contamination of vegetables. Foram analisadas amostras de água de irrigação e vegetais de hortas de Campinas, SP. A análise bacteriana condenou 22,3% das hortaliças e o exame parasitológico condenou 14,5%. O critério estabelecido pela legislação brasileira condenou 11,8% das amostras de água de irrigação. A contaminação de hortaliças por parasitas foi significantemente mais freqüente na estação chuvosa, enquanto o excesso de coliformes fecais foi mais freqüente na estação seca. O monitoramento adequado da água de irrigação é importante para evitar a contaminação de vegetais.

Changes on cyclic adenosine monophosphate (cAMP) levels in response to adenosine and glutamate and the subtype of glutamate receptors involved in this interaction were studied in slices of optic tectum from 3-day-old chicks. cAMP accumulation mediated by adenosine (100 microM) was abolished by 8-phenyltheophylline (15 microM). Glutamate and the glutamatergic agonists kainate or trans-D, L-1-aminocyclopentane-1,3-dicarboxylic acid (trans-ACPD) did not evoke cAMP accumulation. Glutamate blocked the adenosine response in a dose-dependent manner. At 100 microM, glutamate did not inhibit the effect of adenosine. The 1 mM and 10 mM doses of glutamate inhibited adenosine-induced cAMP accumulation by 55% and 100%, respectively. When glutamatergic antagonists were used, this inhibitory effect was not affected by 200 microM 6,7-dihydroxy-2,3,dinitroquinoxaline (DNQX), an ionotropic antagonist, and was partially antagonized by 1 mM (RS)-alpha-methyl-4-carboxyphenylglycine [(RS)M-CPG], a metabotropic antagonist, while 1 mM L-2-amino-3-phosphonopropionate (L-AP3) alone, another metabotropic antagonist, presented the same inhibitory effect of glutamate. Kainate (10 mM) and trans-ACPD (100 microM and 1 mM) partially blocked the adenosine response. This study indicates the involvement of metabotropic glutamate receptors in adenylate cyclase inhibition induced by glutamate and its agonists trans-ACPD and kainate.

This study reports the results of microscopic analysis for detecting and identifying magnetic particles in granulated sugar produced and commercialized in the regions of Ribeirao Preto, Bauru and Campinas of Sao Paulo State. The National Health Surveillance Agency through the REALI (Alert and Communication of Risks in Foods Network) no17/2011 requested for the Sanitary Surveillance Groups action for supervising and collecting samples from sugar factories after consumers complaints. Sixty samples collected between 2011 e 2012 were analyzed at the Centers for Regional Laboratories of these three regions. The methodologies of scattering sample (Zamboni, 1986) and of dissolution of sample (AOAC, 2005) were employed for detecting the magnetic particles, and the results were evaluated according to legislations in force. Of 41 samples collected in 2011, 41.5 % contained magnetic particles up to 4.5 mm of length, and of 19 samples analyzed in 2012, 21.1 % contained particles up to 2.0 mm, which indicate fails in good practical of manufacture and potentials health risks, which require corrective measures in production. It is crucial to institute a legislation to set benchmarks and limits for the presence of these particles, and the greater integration among industry, scientific community and regulatory agencies to ensure safe food production.

The aim with this investigation was to study the aerodynamic properties of truck-trailer combinations of varying lengths. The aerodynamic properties of the combinations were evaluated in order to study similarities and differences in the flow field between different configurations. By the use of Computational Fluid Dynamics (CFD) six different types of truck-trailer combinations used for long hauling have been evaluated. The combinations have a total length varying between 10.10 m and 25.25 m and consist of either a tractor or rigid truck in combination with one or two cargo units. All of the combinations are commonly found on roads in Sweden and several other countries in Europe. The results from the simulations show that the aerodynamic properties differ significantly for the truck-trailer combinations. It was found that the longer vehicle combinations are much more sensitive to yaw conditions than the shorter combinations. What was also evident is that the gap between the first and second cargo unit plays a significant role for the aerodynamic properties. Furthermore it was established that there is potential for fuel saving by a strategic choice of truck-trailer combination. According to the simplified evaluation made in this study the 25.25 m combinations show a lower power required per loading length (LL) and are therefore shown to be more efficient than the shorter combinations.