Rapid detection of bovine coronavirus by a semi-nested RT-PCR

Bovine coronavirus (BCoV) is a member of the group 2 of the Coronavirus (Nidovirales: Coronaviridae) and the causative agent of enteritis in both calves and adult bovine, as well as respiratory disease in calves. The present study aimed to develop a semi-nested RT-PCR for the detection of BCoV based on representative up-to-date sequences of the nucleocapsid gene, a conserved region of coronavirus genome. Three primers were designed, the first round with a 463bp and the second (semi-nested) with a 306bp predicted fragment. The analytical sensitivity was determined by 10-fold serial dilutions of the BCoV Kakegawa strain (HA titre: 256) in DEPC treated ultra-pure water, in fetal bovine serum (FBS) and in a BCoV-free fecal suspension, when positive results were found up to the 10-2, 10-3 and 10-7 dilutions, respectively, which suggests that the total amount of RNA in the sample influence the precipitation of pellets by the method of extraction used. When fecal samples was used, a large quantity of total RNA serves as carrier of BCoV RNA, demonstrating a high analytical sensitivity and lack of possible substances inhibiting the PCR. The final semi-nested RT-PCR protocol was applied to 25 fecal samples from adult cows, previously tested by a nested RT-PCR RdRp used as a reference test, resulting in 20 and 17 positives for the first and second tests, respectively, and a substantial agreement was found by kappa statistics (0.694). The high sensitivity and specificity of the new proposed method and the fact that primers were designed based on current BCoV sequences give basis to a more accurate diagnosis of BCoV-caused diseases, as well as to further insights on protocols for the detection of other Coronavirus representatives of both Animal and Public Health importance.

Factors affecting immunoglobulin concentration in colostrum of healthy Holstein cows immediately after delivery

This study analyzed the influence of the number of milkings, number of births, and udder quarter in immunoglobulin (Ig) concentration in the colostrum of healthy Holstein cows. It was collected two samples of colostrum by manual milking, getting the first jets to completion of bacteriological examination and immunoglobulin levels by radial immunodiffusion test in agar gel. Positive samples for bacteriological examination were excluded from this investigation. Medians of immunoglobulin's G, A and M in the colostrum collected before the first and second milking were respectively 9,200 and 6,400mg/dL (p=0.0029); 400 and 200mg/dL (p=0.0018); 800 and 400mg/dL (p=0.0001). Median immunoglobulin concentration in animals that calved once, twice or three times or in cows that calved 4 to 6 times were 6,400; 6,400; 3,200 and 11,200mg/dL IgG; 100, 200, 100 and 800mg/dL IgA ; and 400, 400, 100 and 800mg/dL IgM, respectively. Concentrations of IgG, IgA and IgM were greater in animals that calved more than 4 times (p<0.05). Medians of IgG, IgA and IgM in the right fore quarter (RF), right hind quarter (RH), left fore quarter (LF) and left hind quarter (LH) were, respectively, 7,800; 6,400; 7,800 and 6,400mg/dL; 200, 200, 200 and 200mg/dL; and 400, 400, 400 and 400mg/dL. Ig concentrations in the colostrum of Holstein cows were influenced by the number of milkings after delivery and number of lactations. These variations may be considered risk factors to passive immunity transfer to newborn calves, predisposing them to diseases and causing economic losses to dairy production.

Effects of weed "Reestablishment" after hoeing on corn yields

Some growers and researchers sustain the idea that regrowth or root setting of some weeds may occur after hoeing, with detrimental effects over corn. The objective of this study was to evaluate the effects of weed removal from the field, removal after each hoeing, and corn intercropped with gliricidia on weed control and corn yield values. The experimental design consisted of blocks with split-plots and six replicates. Cultivars AG 1051 and BM 2022, planted in the plots, were submitted to the following treatments: no hoeing, two hoeings (at 20 and 40 days after planting), and intercropped with gliricidia. The hoed plots were either submitted to weed removal after the first, second, or both hoeings, or remained without weed removal. In the intercropped treatment, gliricidia was sown by broadcasting at corn planting between the corn rows, at a density of 15 seeds m-2. Twenty-five weed species occurred in the experiment; the most frequent was Digitaria sanguinalis (family Poaceae). The weed control methods tested had similar effects on the cultivars, which were not different from one another with respect to the evaluated traits, except for one-hundred-kernel weight, with cultivar AG 1051 being superior. Weed removal did not influence green corn yield or grain yield. However, the number of kernels/ear was higher in plots where weeds were removed in relation to plots without weed removal, suggesting that weed removal might be beneficial to corn. Besides, a higher dry matter weight was obtained for the above-ground part of weeds removed from the field after the first and second hoeings than the weight of weeds removed after the second hoeing only which, in turn, was higher than the weight of weeds removed after the first hoeing only. Green ear yield, grain yield, and dry matter of the above-ground part of the weeds did not show differences in hoed plots and were superior to the non-weeded plots and the intercropped plots, which were not different from each other; therefore, intercropping with gliricidia did not improve corn yield values.

EFFECT OF POST-EMERGENCE HERBICIDE APPLICATIONS ON RICE CROP WEED COMMUNITIES IN TOLIMA, COLOMBIA

ABSTRACTThe weed communities of agricultural systems are dynamic and respond to changes in agronomic practices. This study aimed to evaluate the effect of post-emergence herbicide control programs made by farmers on weed communities and commercial lots of rice. The evaluations were carried out in 96 commercial lots located in the Centro, Meseta and Norte zones of the department of Tolima. In each lot, 1 ha was marked off, in which the evaluations were carried out by randomly throwing a 0.2 x 0.2 m sampling-square 5 times. Samples were taken before the first post-emergence application, after the first post-emergence application, after the second post-emergence application, and once the post-emergence applications were finished. The evaluated variables included density and cover of the weeds and the crops. The IVI of each species was calculated and the control program was analyzed in terms of decreases in the number of individuals for the 15 more encountered species. Before the applications, higher density values were found. The first and second post-emergence applications reduced the average density by 41% and 12%, respectively, throughout the department. Between the first and fourth evaluations, the density of the weeds and crops decreased throughout the department by 51.7% and 39%, respectively. The weed density variable proved to be the most influential in the populations after the herbicide programs were carried out.

Effects of septal cholinergic lesion on rat exploratory behavior in an open-field

The medial septum participates in the modulation of exploratory behavior triggered by novelty. Also, selective lesions of the cholinergic component of the septohippocampal system alter the habituation of rats to an elevated plus-maze without modifying anxiety indices. We investigated the effects of the intraseptal injection of the cholinergic immunotoxin 192 IgG-saporin (SAP) on the behavior of rats in an open-field. Thirty-nine male Wistar rats (weight: 194-230 g) were divided into three groups, non-injected controls and rats injected with either saline (0.5 µl) or SAP (237.5 ng/0.5 µl). Twelve days after surgery, the animals were placed in a square open-field (120 cm) and allowed to freely explore for 5 min. After the test, the rats were killed by decapitation and the septum, hippocampus and frontal cortex were removed and assayed for acetylcholinesterase activity. SAP increased acetylcholinesterase activity in the septum, hippocampus and frontal cortex and decreased the total distance run (9.15 ± 1.51 m) in comparison to controls (13.49 ± 0.91 m). The time spent in the center and at the periphery was not altered by SAP but the distance run was reduced during the first and second minutes (2.43 ± 0.36 and 1.75 ± 0.34 m) compared to controls (4.18 ± 0.26 and 3.14 ± 0.25 m). SAP-treated rats showed decreased but persistent exploration throughout the session. These results suggest that septohippocampal cholinergic mechanisms contribute to at least two critical processes, one related to the motivation to explore new environments and the other to the acquisition and storage of spatial information (i.e., spatial memory).

Opioidergic orofacial antinociception induced by electroacupuncture at acupoint St36

The participation of opioids in the antinociceptive effect of electroacupuncture was evaluated in terms of nociception produced by thermal stimuli applied to the face of male Wistar rats, weighing 180-230 g. Electrical stimulation (bipolar and asymmetric square wave with 0.5 mA intensity for 20 min) of acupoint St36, located in the anterior tibial muscle 10 mm distal to the knee joint, induced antinociception in the present model, which was maintained for 150 min. Acupoint LI4, located in the junction of the first and second metacarpal bones, did not achieve antinociception at any frequency studied (5 Hz: 1.7 ± 0.1; 30 Hz: 1.8 ± 0.1; 100 Hz: 1.7 ± 0.1 vs 1.4 ± 0.2). The antinociception obtained by stimulation of acupoint St36 was only achieved when high frequency 100 Hz (3.0 ± 0.2 vs 1.0 ± 0.1) was used, and not with 5 or 30 Hz (1.2 ± 0.2 and 0.7 ± 0.1 vs 1.0 ± 0.1). The antinociceptive effect of acupuncture occurred by opioid pathway activation, since naloxone (1 and 2 mg/kg, subcutaneously) antagonized it (1.8 ± 0.2 and 1.7 ± 0.2 vs 3.0 ± 0.1).

Phenylpyrazolone derivatives inhibit gastric emptying in rats by a capsaicin-sensitive afferent pathway

Dipyrone (Dp), 4-aminoantipyrine (AA) and antipyrine (At) administered iv and Dp administered icv delay gastric emptying (GE) in rats. The participation of capsaicin (Cps)-sensitive afferent fibers in this phenomenon was evaluated. Male Wistar rats were pretreated sc with Cps (50 mg/kg) or vehicle between the first and second day of life and both groups were submitted to the eye-wiping test. GE was determined in these animals at the age of 8/9 weeks (weight: 200-300 g). Ten minutes before the study, the animals of both groups were treated iv with Dp, AA or At (240 μmol/kg), or saline; or treated icv with Dp (4 μmol/animal) or saline. GE was determined 10 min after treatment by measuring % gastric retention (GR) of saline labeled with phenol red 10 min after orogastric administration. Percent GR (mean ± SEM, N = 8) in animals pretreated with Cps and treated with Dp, AA or At (35.8 ± 3.2, 35.4 ± 2.2, and 35.6 ± 2%, respectively) did not differ from the GR of saline-treated animals pretreated with vehicle (36.8 ± 2.8%) and was significantly lower than in animals pretreated with vehicle and treated with the drugs (52.1 ± 2.8, 66.2 ± 4, and 55.8 ± 3%, respectively). The effect of icv administration of Dp (N = 6) was not modified by pretreatment with Cps (63.3 ± 5.7%) compared to Dp-treated animals pretreated with vehicle (62.3 ± 2.4%). The results suggest the participation of capsaicin-sensitive afferent fibers in the delayed GE induced by iv administration of Dp, AA and At, but not of icv Dp.

Effect of melatonin on the functional recovery from experimental traumatic compression of the spinal cord

Spinal cord injury is an extremely severe condition with no available effective therapies. We examined the effect of melatonin on traumatic compression of the spinal cord. Sixty male adult Wistar rats were divided into three groups: sham-operated animals and animals with 35 and 50% spinal cord compression with a polycarbonate rod spacer. Each group was divided into two subgroups, each receiving an injection of vehicle or melatonin (2.5 mg/kg, intraperitoneal) 5 min prior to and 1, 2, 3, and 4 h after injury. Functional recovery was monitored weekly by the open-field test, the Basso, Beattie and Bresnahan locomotor scale and the inclined plane test. Histological changes of the spinal cord were examined 35 days after injury. Motor scores were progressively lower as spacer size increased according to the motor scale and inclined plane test evaluation at all times of assessment. The results of the two tests were correlated. The open-field test presented similar results with a less pronounced difference between the 35 and 50% compression groups. The injured groups presented functional recovery that was more evident in the first and second weeks. Animals receiving melatonin treatment presented more pronounced functional recovery than vehicle-treated animals as measured by the motor scale or inclined plane. NADPH-d histochemistry revealed integrity of the spinal cord thoracic segment in sham-operated animals and confirmed the severity of the lesion after spinal cord narrowing. The results obtained after experimental compression of the spinal cord support the hypothesis that melatonin may be considered for use in clinical practice because of its protective effect on the secondary wave of neuronal death following the primary wave after spinal cord injury.

Impact of specimen adequacy on the assessment of renal allograft biopsy specimens

The Banff classification was introduced to achieve uniformity in the assessment of renal allograft biopsies. The primary aim of this study was to evaluate the impact of specimen adequacy on the Banff classification. All renal allograft biopsies obtained between July 2010 and June 2012 for suspicion of acute rejection were included. Pre-biopsy clinical data on suspected diagnosis and time from renal transplantation were provided to a nephropathologist who was blinded to the original pathological report. Second pathological readings were compared with the original to assess agreement stratified by specimen adequacy. Cohen's kappa test and Fisher's exact test were used for statistical analyses. Forty-nine specimens were reviewed. Among these specimens, 81.6% were classified as adequate, 6.12% as minimal, and 12.24% as unsatisfactory. The agreement analysis among the first and second readings revealed a kappa value of 0.97. Full agreement between readings was found in 75% of the adequate specimens, 66.7 and 50% for minimal and unsatisfactory specimens, respectively. There was no agreement between readings in 5% of the adequate specimens and 16.7% of the unsatisfactory specimens. For the entire sample full agreement was found in 71.4%, partial agreement in 20.4% and no agreement in 8.2% of the specimens. Statistical analysis using Fisher's exact test yielded a P value above 0.25 showing that - probably due to small sample size - the results were not statistically significant. Specimen adequacy may be a determinant of a diagnostic agreement in renal allograft specimen assessment. While additional studies including larger case numbers are required to further delineate the impact of specimen adequacy on the reliability of histopathological assessments, specimen quality must be considered during clinical decision making while dealing with biopsy reports based on minimal or unsatisfactory specimens.

Edistyneen oppijan suomen konstruktiopiirteitä korpusvetoisesti: avainrakenneanalyysi

Ensikielen jälkeen opittavan kielen tutkimusta ja suomi toisena kielenä alaa sen osana ovat koko niiden olemassaolon ajan hallinneet samat peruskysymykset: millaista oppiminen on eri vaiheissa ja eri ympäristöissä, sekä mikä oppimisessa on yleistä ja toisaalta mikä riippuu opittavasta kielestä ja oppijoiden kielitaustasta. Sähköisten oppijankielen tutkimusaineistojen eli korpusten lisääntymisen myötä tutkijat voivat aiempaa helpommin tutkia näitä ilmiöitä määrällisesti ja tarkastella oppijankielen sisäistä vaihtelua ja sen suhdetta tyypilliseen ensikieliseen kielenkäyttöön kielen eri osa-alueilla käyttöpohjaisesti eli todelliseen kielenkäyttöön pohjautuen. Tekninen kehitys on tuonut mukanaan aineisto- eli korpusvetoisuuden kaltaisia uusia tapoja lähestyä tutkimusaineistoa, jolloin tyypillisiä tutkimuskysymyksiä ”Miksi?” ja ”Miten?” edeltää kysymys: ”Mikä?”. Tässä väitöskirjassa tarkastellaan edistyneiden suomenoppijoiden kirjoitettua akateemista kieltä ja suhteutetaan suomen oppimiselle ominaisia seikkoja käyttöpohjaisen mallin perusolettamuksiin. Aineisto on suomea toisena kielenä käyttävien opiskelijoiden tenttivastauksia, ja se on osa Edistyneiden suomenoppijoiden korpusta. Tutkimus on osin metodologinen, sillä väitöskirjassa esitellään ja siinä sovelletaan uutta korpusvetoista avainrakenneanalyysi-menetelmää, jonka avulla aineistoa lähestytään ilman hypoteeseja siitä, mitkä kielen ilmiöt ovat ominaisia edistyneelle oppijansuomelle. Tutkimus kuuluu kieliopin tutkimuksen piiriin, ja se nojaa kognitiivisen konstruktiokieliopin ajatukseen abstraktiudeltaan vaihtelevista konstruktioista kielijärjestelmän perusyksiköinä. Tulokset puoltavat menetelmän sovellettavuutta kielen oppimisen tutkimukseen, sillä sen avulla kyettiin tunnistamaan konstruktioita, jotka erottavat edistyneitä oppijoita ensikielisistä kirjoittajista (esim. modaaliset verbiketjut), eri ensikieliä puhuvia suomenoppijoita (esim. konjunktiot) sekä konstruktioita, joiden käyttö muuttuu ajan kuluessa (esim. preteriti ja preesens). Monet havaitut erot ovat akateemisen kirjoittamisen erityispiirteitä, mikä tukee ajatusta kielen käyttö- ja kontekstikohtaisesta oppimisesta. Tuloksia voidaan yhtäältä soveltaa akateemisen kielitaidon opetuksessa. Toisaalta menetelmää voidaan käyttää kielenoppimisen tutkimuksen ohella uusien näkökulmien kartoittamiseksi erilaisten tai eri-ikäisten tekstien tyypillisten ominaisuuksien ja erojen tutkimuksessa.

Optimal processing chain for waste power plant

This study is done to examine waste power plant’s optimal processing chain and it is important to consider from several points of view on why one option is better than the other. This is to insure that the right decision is made. Incineration of waste has devel-oped to be one decent option for waste disposal. There are several legislation matters and technical options to consider when starting up a waste power plant. From the tech-niques pretreatment, burner and flue gas cleaning are the biggest ones to consider. The treatment of incineration residues is important since it can be very harmful for the envi-ronment. The actual energy production from waste is not highly efficient and there are several harmful compounds emitted. Recycling of waste before incineration is not very typical and there are not many recycling options for materials that cannot be easily re-cycled to same product. Life cycle assessment is a good option for studying the envi-ronmental effect of the system. It has four phases that are part of the iterative study process. In this study the case environment is a waste power plant. The modeling of the plant is done with GaBi 6 software and the scope is from gate-to-grave. There are three different scenarios, from which the first and second are compared to each other to reach conclusions. Zero scenario is part of the study to demonstrate situation without the power plant. The power plant in this study is recycling some materials in scenario one and in scenario two even more materials and utilize the bottom ash more ways than one. The model has the substitutive processes for the materials when they are not recycled in the plant. The global warming potential results show that scenario one is the best option. The variable costs that have been considered tell the same result. The conclusion is that the waste power plant should not recycle more and utilize bottom ash in a number of ways. The area is not ready for that kind of utilization and production from recycled materials.

Reconstruction of cranial bone defects with fiber-reinforced composite–bioactive glass implants

A cranial bone defect may result after an operative treatment of trauma, infection, vascular insult, or tumor. New biomaterials for cranial bone defect reconstructions are needed for example to mimic the biomechanical properties and structure of cranial bone. A novel glass fiber-reinforced composite implant with bioactive glass particulates (FRC–BG, fiber-reinforced composite–bioactive glass) has osteointegrative potential in a preclinical setting. The aim of the first and second study was to investigate the functionality of a FRC–BG implant in the reconstruction of cranial bone defects. During the years 2007–2014, a prospective clinical trial was conducted in two tertiary level academic institutions (Turku University Hospital and Oulu University Hospital) to evaluate the treatment outcome in 35 patients that underwent a FRC–BG cranioplasty. The treatment outcome was good both in adult and pediatric patients. A number of conventional complications related to cranioplasty were observed. In the third study, a retrospective outcome evaluation of 100 cranioplasty procedures performed in Turku University Hospital between years 2002–2012 was conducted. The experimental fourth study was conducted to test the load-bearing capacity and fracture behavior of FRC–BG implants under static loading. The interconnective bars in the implant structure markedly increased the load-bearing capacity of the implant. A loading test did not demonstrate any protrusions of glass fibers or fiber cut. The fracture type was buckling and delamination. In this study, a postoperative complication requiring a reoperation or removal of the cranioplasty material was observed in one out of five cranioplasty patients. The treatment outcomes of cranioplasty performed with different synthetic materials did not show significant difference when compared with autograft. The FRC–BG implant was demonstrated to be safe and biocompatible biomaterial for large cranial bone defect reconstructions in adult and pediatric patients.

Physiological response of cowpea seeds to salinity stress

This work aimed to evoluate physiological response of cowpea (Vigna unguiculata L. Walp) seeds submitted to salt stress. Seeds of cultivars 'Epace-10'; 'Canapu' and 'Pitiúba' of cowpea, were submitted to germination test in germinator at 25(0)C, in "germitest" papers imbibed in distilled water or in 0, 10, 50, 100 and 200mol m-3 NaCl solutions. At the first and second counting of the germination test, normal seedlings were accounted, weighted and dried, obtaining data for vigor, total germination, fresh matter weight and dry matter weight. The seedlings hypocotyls, root and total length were measured total proteins content in cotyledons were obtained from germinating seeds. The presence of salt at concentrations higher than 50mol m-3 NaCl affect the germination, seedlings growth and cotyledons total protein synthesis of all cowpea cultivars. The seeds of cultivar pitiúba were is more tolerant to salinity, than the cultivars Canapu and Epace-10.

Grammatikdidaktik: Svensk grammatik och språklig medvetenhet i några läromedel för grundskolans första årskurser

Mitt övergripande forskningsintresse är grammatikdidaktik, medan det explicita syftet med denna licentiatavhandling är att erhålla ny kunskap om grammatikdidaktik så som den synliggörs i nutida läromedel för grundskolans första årskurser. Den empiriska undersökningen är gjord i tre nutida läromedelspaket avsedda för undervisning i skolämnet modersmål och litteratur i årskurs ett och två, läromedlen som undersöks är de finlandssvenska serierna Läseboken och Litteraturboken samt den sverigesvenska serien Kom och läs! Bland resultaten kan nämnas att det finns ett brett spektrum av nivåer av extern grammatik i alla de undersökta läromedelspaketen. Den fonologiska och syntaktiska medvetenheten, vilka bägge är viktiga för läs- och skrivutvecklingen i de diskuterade årskurserna, satsas på i alla tre läromedelspaket. Det jag dock också märkt i läromedlen som kan påverka läs- och skrivutvecklingen är att andelen belägg för grafonomi är större i årskurs två än i årskurs ett. Härigenom får eleverna på detta sätt småningom växa in i en skriftkultur. Det finns betydligt fler muntliga uppgifter i lärarhandledningarna till serierna Litteraturboken och Kom och läs! än i Läseboken. Det exakta ”hur” står inte alltid utskrivet i lärarhandledningarna, men de flesta gånger verkar tanken vara att det rör sig om diskussioner eller andra muntliga genrer. I många fall känns de muntliga uppgifterna mer funktionella än skriftliga uppgifter som inte verkar vara helt genomtänkta. Att mitt material innehåller fler funktionella uppgifter i lärarhandledningar och fler formalistiska eller delvis funktionella uppgifter i arbetsböcker överensstämmer med att lärarhandledningar kan vara en bättre väg för att öka elevernas kunskapssökande och minska förmedlingspedagogiken.

An examination of the effects on learning seen in children afforded the opportunity to control the order of repetitions for three novel spatiotemporal sequences

Children were afforded the opportunity to control the order of repetitions for three novel spatiotemporal sequences. The following was predicted: a) children and adults in the self-regulated (SELF) groups would produce faster movement (MT) and reaction times (R T) and greater recall success (RS) during retention compared to the age-matched yoked (YOKE) groups; b) children would choose to switch sequences less often than adults; c) adults would produce faster MT and RT and greater RS than the children during acquisition and retention, independent of experimental group. During acquisition, no effects were seen for RS, however for MT and RT there was a main effect for age as well as block. During retention a main effect for practice condition was seen for RS and failed to reach statistical significance for MT and RT, thus partially supporting our first and second hypotheses. The third hypothesis was not supported.