Development and validation of a method for detection and quantification of ochratoxin A in green coffee using liquid chromatography coupled to mass spectrometry

A method using Liquid Chromatography Tanden Mass Spectrometry (LC-MS/MS) with matrix-matched calibration curve was developed and validated for determining ochratoxin A (OTA) in green coffee. Linearity was found between 3.0 and 23.0 ng.g-1. Mean recoveries ranged between 90.45% and 108.81%; the relative standard deviation under repeatability and intermediate precision conditions ranged from 5.39% to 9.94% and from 2.20% to 14.34%, respectively. The limits of detection and quantification were 1.2 ng.g-1 and 3.0 ng.g-¹, respectively. The method developed was suitable and contributed to the field of mycotoxin analysis, and it will be used for future production of the Certified Reference Material (CRM) for OTA in coffee.

Assessment of toxic potential of Cerrado fruit seeds using Artemia salina bioassay

Artemia salina bioassay was used to assess toxicity of seeds and kernels of Brazilian fruits from cerrado (central high plains region) and other inner regions of the country. Water extracts of the kernels were filtered and added to Artemia cultures containing ten individuals per mL. Dose - response curves were constructed, and LD50 values were calculated. Pure potassium cyanide standard was used to draw a calibration curve for comparison to detect the presence of cyanide in the samples tested. Extracts of the seeds of araticum, mangaba, cagaita, jatobá, and tucumã were found toxic to Artemia salina, and some of the dose - response curves were very similar in shape to those obtained with pure potassium cyanide standards, while the samples of baru, cajá-manga, siriguela, trauma, and veludo showed no toxicity at all. The Guignard test, specific for cyanide detection, showed negative results in all toxic samples, suggesting the presence of other toxic compounds rather than cyanide. The comparison of araticum dose - response curve with those of other annonaceous fruits suggests the presence acetogenins as the main toxic compounds in the seeds. These results could be useful to prevent poisoning by industrial derivatives of the fruits studied.

Forced-air, vacuum, and hydro precooling of cauliflower (Brassica oleracea L. var. botrytis cv. Freemont): part I. determination of precooling parameters

The aim of the present study was to precool cauliflower using forced-air, vacuum and high and low flow hydro cooling methods. The weight of the precooled cauliflower heads (5000±5 g) was measured before they were placed in standard plastic crates. Cauliflower heads, whose initial temperature was 23.5 ± 0.5 ºC, were cooled until the temperature reached at 1 ºC. During the precooling process, time-dependent temperature and energy consumption were measured, and during vacuum precooling, the decreasing pressure values were recorded, and a curve of time-dependent pressure decrease (vacuum) was built. The most suitable cooling method to precool cauliflower in terms of cooling time and energy consumption was vacuum, followed by the high and low flow hydro and forced-air precooling methods, respectively. The highest weight loss was observed in the vacuum precooling method, followed by the forced-air method. However, there was an increase in the weight of the cauliflower heads in the high and low flow hydro precooling method. The best colour and hardness values were found in the vacuum precooling method. Among all methods tested, the most suitable method to precool cauliflower in terms of cooling and quality parameters was the vacuum precooling method.

Effect of standard and neutral-pH peritoneal dialysis solutions upon fibroblasts proliferation

Introduction: Continuous exposition of the peritoneal membrane to conventional dialysis solutions is an important risk factor for inducing structural and functional alterations. Objective: To compare in vitro mouse fibroblast NIH-3T3 cell viability after exposition to a neutral pH dialysis solution in comparison to cells exposed to a standard solution. Methods: Experimental study to compare the effects of a conventional standard or a neutral-pH, low-glucose degradation products peritoneal dialysis solution on the viability of exposed fibroblasts in cell culture. Both solutions were tested in all the commercially available glucose concentrations. Cell viability was evaluated with tetrazolium salt colorimetric assay. Results: Fibroblast viability was significantly superior in the neutral pH solution in comparison to control, in all three glucose concentrations (Optical density in nm-means ± SD: 1.5% 0.295 ± 0.047 vs. 0.372 ± 0.042, p < 0.001; 2.3% 0.270 ± 0.036 vs. 0.337 ± 0.051, p < 0.001; 4.25% 0.284 ± 0.037 vs. 0.332 ± 0.032, p < 0.001; control vs. neutral pH respectively, Student t Test). There was no significant difference in cell viability between the three concentrations of glucose when standard solution was used (ANOVA p = 0.218), although cell viability was higher after exposition to neutral pH peritoneal dialysis fluid at 1.5% in comparison to 2.3 and 4.25% glucose concentrations (ANOVA p = 0.008: Bonferroni 1.5% vs. 2.3% p = 0.033, 1.5% vs. 4.25% p = 0.014, 2.3% vs. 4.25% p = 1.00). Conclusion: Cell viability was better in neutral pH dialysis solution, especially in the lower glucose concentration. A more physiological pH and lower glucose degradation products may be responsible for such results.

Analysis and guidelines for implementation of EN 1090-2 Standard to assist execution class assignment of steel structures

Finnish design and consulting companies are delivering robust and cost-efficient steel structures solutions to a large number of manufacturing companies worldwide. Recently introduced EN 1090-2 standard obliges these companies to specify the execution class of steel structures for their customers. This however, requires clarifying, understanding and interpreting the sophisticated procedure of execution class assignment. The objective of this research is to provide a clear explanation and guidance through the process of execution class assignment for a given steel structure and to support the implementation of EN 1090-2 standard in Rejlers Oy, one of Finnish design and consulting companies. This objective is accomplished by creating a guideline for designers that elaborates on the four-step process of the execution class assignment for a steel structure or its part. Steps one to three define the consequence class (projected consequences of structure failure), the service category (hazards associated with the service use exploitation of steel structure) and the production category (manufacturing process peculiarities), based on the ductility class (capacity of structure to withstand deformations) and the behaviour factor (corresponds to structure seismic behaviour). The final step is the execution class assignment taking into account results of previous steps. Main research method is indepth literature review of European standards family for steel structures. Other research approach is a series of interviews of Rejlers Oy representatives and its clients, results of which have been used to evaluate the level of EN 1090-2 awareness. Rejlers Oy will use the developed novel coherent standard implementation guideline to improve its services and to obtain greater customer satisfaction.

Marks of Fire, Value and Faith. Swords with Ferrous Inlays in Finland during the Late Iron Age (ca. 700–1200 AD)

This study explores swords with ferrous inlays found in Finland and dating from the late Iron Age, ca. 700–1200 AD. These swords reflect profound changes not only in styles and fashion but also in the technology of hilts and blades. This study explores how many of these kinds of swords are known from Finland, how they were made and where, what their status was in Late Iron Age Finland, and where the Finnish finds stand in accordance with other areas of Europe. The various methods included measuring of the finds and statistics. The main method of revealing the inlaid marks was radiography due to its non-destructive nature. In cases where inlays were visible without radiography, their details were inspected via microscopy. To study the materials and manufacture of inlaid swords, a sample of them was metallographically analysed to determine the forging technologies and nature of used materials. Furthermore, the manufacture was also studied with experimental approaches. As a result, a catalogue of 151 swords with ferrous inlays was created. This number is relatively high compared with other European countries, although systematic studies have been conducted in only some countries. The inlaid motifs were classified into five distinct categories to help the classification. To summarize, almost every documented inlaid sword was unique in some respect including measurements, inlaid motifs and materials of blades and inlays. Technological variation was also present, some blades being poorer and some of higher quality in spite of the inlaid motifs. Misspelt inscriptions as well as letter-like marks were common in Finland and also in Scandinavia. Furthermore, the provenance of iron and steel used in some blades hints at Scandinavian ores. The above observations, along with the experimental results indicating the existence of multiple alternative techniques of inlaying, suggest that these swords were manufactured locally in Scandinavia, most likely in imitation of Continental European models. Inlaid swords were valued partly for their assumed functionality in combat, as evidenced by damage on some examined blades, or they were valued for their inlays, which could have had fashionable or symbolical meanings bound to local beliefs.

Pienten yhtiöiden tilintarkastusstandardin arvioitu vaikutus tilintarkastuksen hyötyihin – Standard for Audits of Small Entities, SASE

Suomessa ja muissa Pohjoismaissa on tunnistettu haasteita kansainvälisten tilintarkastusstandardien eli ISA-standardien soveltamisessa pienten yhtiöiden tilintarkastuksiin. ISA-standardien on kritisoitu sisältävän liian yksityiskohtaisia ja jopa epärealistisia vaatimuksia ajatellen pienien yhtiöiden tilintarkastuksia. Pohjoismaisella yhteistyöllä onkin alettu kehittää omaa standardia pienten yhtiöiden tilintarkastuksiin. Niin sanotun SASE-standardin on raportoitu muun muassa korostavan tilintarkastajan ammatillista harkintaa ja keventävän pienten yhtiöiden tilintarkastuksen vaatimuksia. Koska kuitenkin aikaisemmin sääntelyn lisäämisellä on pyritty muun muassa lisääntyneeseen tilintarkastuksen luotettavuuteen, josta myös pienten yhtiöiden on itse tutkittu hyötyvän, on tutkimuksessa haluttu selvittää, miten kehitteillä olevan SASE-standardin arvioidaan vaikuttavan yleisesti pienten yhtiöiden tilintarkastuksella saavutettaviin hyötyihin. Tutkimus on toteutettu toiminta-analyyttisena laadullisena tutkimuksena, ja tutkimusmetodina on käytetty tilintarkastajien ja yliopiston tutkijatohtorin haastatteluja. Tutkimuksen tulosten mukaan kehitteillä olevan SASE-standardin arvioidaan vaikuttavan pienten yhtiöiden tilintarkastuksen hyötyihin kaksisuuntaisesti. Toisaalta SASE-standardi voi lisätä tilintarkastuksen hyötyjä, koska tilintarkastaja voisi keskittyä tarkastuskohteen kannalta olennaisempiin asioihin, hyödyntää hiljaista tietoaan sekä lisätä vuorovaikutusta yhtiön omistajajohdon kanssa. SASE-standardin ei arvioida muuttavan juurikaan käytännön tilintarkastustyötä ja periaatteessa olennaisuuksiin keskittymisen ja riskien tehokkaamman tunnistamisen tulisi hyödyttää myös tilintarkastuskohteen ulkoisia sidosryhmiä, kuten rahoittajia. Kuitenkin tutkimuksessa nousi esiin SASE-standardin mahdollinen heikko maine. Mikäli SASE-standardia pidettäisiin vähäpätöisempänä tai epäluotettavampana kuin aikaisempaa käytäntöä, luottamus tilintarkastusta ja tarkastuskohdetta kohtaan heikkenisi ja sitä kautta myös tilintarkastuksen hyödyt pienenisivät. Tutkimuksen johtopäätöksenä voidaan esittää, että tilintarkastuksen sääntelyn keventämisen vaikutus tilintarkastuksen hyötyihin ei niinkään synny käytännön työn kautta. Tilintarkastuksen hyötyihin pikemminkin vaikuttavat maineeseen, viestintään ja vuorovaikutukseen liittyvät seikat. Toimivalla vuorovaikutuksella yhtiön omistajajohdon ja tilintarkastajan välillä voidaan tilintarkastuksen hyötyjä korostaa, ja toisaalta taas riittävällä ulkoisiin sidosryhmiin kohdistuvalla tiedottamisella voidaan hyötyjen heikkenemistä ehkäistä. Keskeiseksi seikaksi tutkimuksessa nousee se, että tilintarkastuksen luotettavuuden ja tason säilyminen on pystyttävä todistamaan.

Interoperability for IoT Health Systems Using HL7 Standard

Internet of Things or IoT is revolutionizing the world we are living in, similarly the way Internet and the web did few decades ago. It is changing how we interact with the things surrounding us. Electronic health and remote patient monitoring are the ways of utilizing these technological improvements towards the healthcare. There are many applications of IoT in eHealth such as, it will open the gate to provide healthcare to the remote areas of the world, where healthcare through traditional hospital systems cannot be provided. To connect these new eHealth IoT systems with the existing healthcare information systems, we can use the existing interoperability standards commonly used in healthcare information systems. In this thesis we implemented an eHealth IoT system based on Health Level 7 interoperability standard for continuous data transmission. There is not much previous work done in implementing the HL7 for continuous sensor data transmission. Some of the previous work was limited to sensors which are not continuous in nature and some of it is only theatrical architecture. This thesis aims to prove that it is possible to implement an eHealth IoT system by using sensors which require continues data transmission, such as respiratory sensors, and to connect it with the existing eHealth information system semantically by using HL7 interoperability standard. This system will be beneficial in implementing eHealth IoT systems for those patients, who requires continuous healthcare personal monitoring. This includes elderly people and patients, whose health need to be monitored constantly. To implement the architecture, HL7 v2.5 is selected due to its ease of implementation and low size. We selected some open source technologies because of their open licenses and large developer community. We will also review the most efficient technology available in every layer of eHealth IoT system and will propose an efficient system.

Development of a bacteriophage-based biopesticide for fire blight

Fire blight is an economically important disease of apples and pears that is caused by the bacterium Erwinia amylovora. Control of the disease depends on limiting primaly blosson1 infection in the spring, and rapidly removing infected tissue. The possibility of using phages to control E.amylovora populations has been suggested, but previous studies have. failed to show high treatment efficacies. This work describes the development of a phage-based biopesticide that controls E. amylovora populations under field conditions, and significantly reduces the incidence of fire blight. This work reports the first use ofPantoea agglomerans, a non-pathogenic relative ofE. amylovora, as a carrier for E. amylovora.phages. Its role is to support a replicating population of these phages on blossom surfaces during the period when the flowers are most susceptible to infection. Seven phages and one carrier isolate were selected for field trials from existing collections of 56 E. amylovora phages and 249 epiphytic orchard bacteria. Selection of the . /' phages and carrier was based on characteristics relevant to the production and field perfonnance of a biopesticide: host range, genetic diversity, growth under the conditions of large-scale production, and the ability to prevent E. amylovora from infecting pear blossoms. In planta assays showed that both the phages and the carrier make significant contributions to reducirig the development of fire blight symptoms in pear blossoms. Field-scale phage production and purification methods were developed based on the growth characteristics of the phages and bacteria in liquid culture, and on the survival of phages in various liquid media. Six of twelve phage-carrier biopesticide treatments caused statistically signiflcant reductions in disease incidence during orchard trials. Multiplex real-time PCR was used to simultaneously monitor the phage, carrier, and pathogen populations over the course of selected treatments. In all cases. the observed population dynamics of the biocontrol agents and the pathogen were consistent with the success or failure of each treatment to control disease incidence. In treatments exhibiting a significantly reduced incidel1ce of fire blight, the average blossom population ofE.amylovora had been reduced to pre-experiment epiphytic levels. In successful treatments the phages grew on the P. agglomerans carrier for 2 to 3 d after treatment application. The phages then grew preferentially on the pathogen, once it was introduced into this blossom ecosystem. The efficacy of the successful phage-based treatnlents was statistically similar to that of streptomycin, which is the most effective bactericide currently available for fire blight prevention. The in planta behaviour ofE. amylovora was compared to that ofErwinia pyrifoliae, a closely related species that causes fire blight-like synlptoms on pears in southeast Asia. Duplex real-time PCR was used to monitor the population dynamics of both species on single blossonls. E. amylovora exhibited a greater competitive fitness on Bartlett pear blossoms than E. pyrifoliae. The genome ofErwinia phage Ea21-4 was sequenced and annotated. Most of the 8-4.7 kB genome is substantially different from previously described sequences, though some regions are notably similar to Salmonella phage Felix 01 . Putative functions were assigned to approximately 30% of the predicted open reading frames based on amino acid sequence comparisons and N-terminal sequencing of structural proteins.

Armourstone revetments : will standard design criteria prevent failure? /

Bank stabilization structures are used to prevent the loss of valuable land within the urban environment and the decision for the type of structure used depends on the properties of the stream. In the urban areas of Southern Ontario there is a preference for the use of armourstone blocks as bank stabilization. The armourstone revetment is a free standing stone structure with large blocks of stone layered vertically and offset from one another. During fieldwork at Forty Mile Creek in Grimsby, Ontario armourstone failure was identified by the removal of two stones within one column from the wall. Since the footer stones were still in place, toe scour was eliminated as a cause of failure. Through theoretical, field, and experimental work the process of suction has been identified as a mode of failure for the armourstone wall and the process of suction works similarly to quarrying large blocks of rock off bedrock streambeds. The theory of lateral suction has previously not been taken into consideration for the design of these walls. The physical and hydraulic evidence found in the field and studied during experimental work indicate that the armourstone wall is vulnerable to the process of suction. The forces exerted by the flow and the resistance of the block determine the stability of the armourstone block within the wall. The design of the armourstone wall, high surface velocities, and short pulses of faster flowing water within the profile could contribute to armourstone failure by providing the forces needed for suction to occur, therefore adjustments to the design of the wall should be made in order to limit the effect.

The role of intrapartum intravenous therapy and method of delivery on newborn weight loss : challenging the 7% rule

It is common practice to initiate supplemental feeding in newborns if body weight decreases by 7-10% in the first few days after birth (7-10% rule). Standard hospital procedure is to initiate intravenous therapy once a woman is admitted to give birth. However, little is known about the relationship between intrapartum intravenous therapy and the amount of weight loss in the newborn. The present research was undertaken in order to determine what factors contribute to weight loss in a newborn, and to examine the relationship between the practice of intravenous intrapartum therapy and the extent of weight loss post-birth. Using a cross-sectional design with a systematic random sample of 100 mother-baby dyads, we examined properties of delivery that have the potential to impact weight loss in the newborn, including method of delivery, parity, duration of labour, volume of intravenous therapy, feeding method, and birth attendant. This study indicated that the volume of intravenous therapy and method of delivery are significant predictors of weight loss in the newborn (R2=15.5, p<0.01). ROC curve analysis identified an intravenous volume cut-point of 1225 ml that would elicit a high measure of sensitivity (91.3%), and demonstrated significant Kappa agreement (p<0.01) with excess newborn weight loss. It was concluded that infusion of intravenous therapy and natural birth delivery are discriminant factors that influence excess weight loss in newborn infants. Acknowledgement of these factors should be considered in clinical practice.