Enhancing performance of paper coatings by tailor-made plastic pigments

The paper industry is constantly looking for new ideas for improving paper products while competition and raw material prices are increasing. Many paper products are pigment coated. Coating layer is the top layer of paper, thus by modifying coating pigment also the paper itself can be altered and value added to the final product. In this thesis, synthesis of new plastic and hybrid pigments and their performance in paper and paperboard coating is reported. Two types of plastic pigments were studied: core-shell latexes and solid beads of maleimide copolymers. Core-shell latexes with partially crosslinked hydrophilic polymer core of poly(n-butyl acrylate-co-methacrylic acid) and a hard hydrophobic polystyrene shell were prepared to improve the optical properties of coated paper. In addition, the effect of different crosslinkers was analyzed and the best overall performance was achieved by the use of ethylene glycol dimethacrylate (EGDMA). Furthermore, the possibility to modify core-shell latex was investigated by introducing a new polymerizable optical brightening agent, 1-[(4-vinylphenoxy)methyl]-4-(2-henylethylenyl)benzene which gave promising results. The prepared core-shell latex pigments performed smoothly also in pilot coating and printing trials. The results demonstrated that by optimizing polymer composition, the optical and surface properties of coated paper can be significantly enhanced. The optimal reaction conditions were established for thermal imidization of poly(styrene-co-maleimide) (SMI) and poly(octadecene-co-maleimide) (OMI) from respective maleic anhydride copolymer precursors and ammonia in a solvent free process. The obtained aqueous dispersions of nanoparticle copolymers exhibited glass transition temperatures (Tg) between 140-170ºC and particle sizes from 50-230 nm. Furthermore, the maleimide copolymers were evaluated in paperboard coating as additional pigments. The maleimide copolymer nanoparticles were partly imbedded into the porous coating structure and therefore the full potential of optical property enhancement for paperboard was not achieved by this method. The possibility to modify maleimide copolymers was also studied. Modifications were carried out via N-substitution by replacing part of the ammonia in the imidization reaction with amines, such as triacetonediamine (TAD), aspartic acid (ASP) and fluorinated amines (2,2,2- trifluoroethylamine, TFEA and 2,2,3,3,4,4,4-heptafluorobuthylamine, HFBA). The obtained functional nanoparticles varied in size between 50-217 nm and their Tg from 150-180ºC. During the coating process the produced plastic pigments exhibited good runnability. No significant improvements were achieved in light stability with TAD modified copolymers whereas nanoparticles modified with aspartic acid and those containing fluorinated groups showed the desired changes in surface properties of the coated paperboard. Finally, reports on preliminary studies with organic-inorganic hybrids are presented. The hybrids prepared by an in situ polymerization reaction consisted of 30 wt% poly(styrene- co-maleimide) (SMI) and high levels of 70 wt% inorganic components of kaolin and/or alumina trihydrate. Scanning Electron Microscopy (SEM) images and characterization by Fourier Transform Infrared Spcetroscopy (FTIR) and X-Ray Diffraction (XRD) revealed that the hybrids had conventional composite structure and inorganic components were covered with precipitated SMI nanoparticles attached to the surface via hydrogen bonding. In paper coating, the hybrids had a beneficial effect on increasing gloss levels.

Chlorophyll fluorescence of tropical tree species in a semi-deciduous forest gap

The characterization of different ecological groups in a forest formation/succession is unclear. To better define the different successional classes, we have to consider ecophysiological aspects, such as the capacity to use or dissipate the light energy available. The main objective of this work was to assess the chlorophyll fluorescence emission of tropical tree species growing in a gap of a semi-deciduous forest. Three species of different ecological groups were selected: Croton floribundus Spreng. (pioneer, P), Astronium graveolens Jacq. (early secondary, Si), and Esenbeckia febrifuga A. Juss. (late secondary, St). The potential (Fv/Fm) and effective (deltaF/Fm') quantum efficiency of photosystem II, apparent electron transport rate (ETR), non-photochemical (qN) and photochemical (qP) quenching of fluorescence were evaluated, using a modulated fluorometer, between 7:30 and 11:00 h. Values of Fv/Fm remained constant in St, decreasing in P and Si after 9:30 h, indicating the occurrence of photoinhibition. Concerning the measurements taken under light conditions (deltaF/Fm', ETR, qP and qN), P and Si showed better photochemical performance, i.e., values of deltaF/Fm', ETR and qP were higher than St when light intensity was increased. Values of qN indicated that P and Si had an increasing tendency of dissipating the excess of energy absorbed by the leaf, whereas the opposite was found for St. The principal component analysis (PCA), considering all evaluated parameters, showed a clear distinction between St, P and Si, with P and Si being closer. The PCA results suggest that chlorophyll fluorescence may be a potential tool to differentiate tree species from distinct successional groups.

“IT’S ALL RIGHT”. Multimodal rightward spatial bias modified by age and praxis

The general goal of the present work was to study whether spatial perceptual asymmetry initially observed in linguistic dichotic listening studies is related to the linguistic nature of the stimuli and/or is modality-specific, as well as to investigate whether the spatial perceptual/attentional asymmetry changes as a function of age and sensory deficit via praxis. Several dichotic listening studies with linguistic stimuli have shown that the inherent perceptual right ear advantage (REA), which presumably results from the left lateralized linguistic functions (bottom-up processes), can be modified with executive functions (top-down control). Executive functions mature slowly during childhood, are well developed in adulthood, and decline as a function of ageing. In Study I, the purpose was to investigate with a cross-sectional experiment from a lifespan perspective the age-related changes in top-down control of REA for linguistic stimuli in dichotic listening with a forced-attention paradigm (DL). In Study II, the aim was to determine whether the REA is linguistic-stimulus-specific or not, and whether the lifespan changes in perceptual asymmetry observed in dichotic listening would exist also in auditory spatial attention tasks that put load on attentional control. In Study III, using visual spatial attention tasks, mimicking the auditory tasks applied in Study II, it was investigated whether or not the stimulus-non-specific rightward spatial bias found in auditory modality is a multimodal phenomenon. Finally, as it has been suggested that the absence of visual input in blind participants leads to improved auditory spatial perceptual and cognitive skills, the aim in Study IV was to determine, whether blindness modifies the ear advantage in DL. Altogether 180-190 right-handed participants between 5 and 79 years of age were studied in Studies I to III, and in Study IV the performance of 14 blind individuals was compared with that of 129 normally sighted individuals. The results showed that only rightward spatial bias was observed in tasks with intensive attentional load, independent of the type of stimuli (linguistic vs. non-linguistic) or the modality (auditory vs. visual). This multimodal rightward spatial bias probably results from a complex interaction of asymmetrical perceptual, attentional, and/or motor mechanisms. Most importantly, the strength of the rightward spatial bias changed as a function of age and augmented praxis due to sensory deficit. The efficiency of the performance in spatial attention tasks and the ability to overcome the rightward spatial bias increased during childhood, was at its best in young adulthood, and decreased as a function of ageing. Between the ages of 5 and 11 years probably at first develops movement and impulse control, followed by the gradual development of abilities to inhibit distractions and disengage attention. The errors especially in bilateral stimulus conditions suggest that a mild phenomenon resembling extinction can be observed throughout the lifespan, but especially the ability to distribute attention to multiple targets simultaneously decreases in the course of ageing. Blindness enhances the processing of auditory bilateral linguistic stimuli, the ability to overcome a stimulus-driven laterality effect related to speech sound perception, and the ability to direct attention to an appropriate spatial location. It was concluded that the ability to voluntarily suppress and inhibit the multimodal rightward spatial bias changes as a function of age and praxis due to sensory deficit and probably reflects the developmental level of executive functions.

Predictors of "occult" intra-abdominal injuries in blunt trauma patients

Objective: to assess predictors of intra-abdominal injuries in blunt trauma patients admitted without abdominal pain or abnormalities on the abdomen physical examination. Methods: We conducted a retrospective analysis of trauma registry data, including adult blunt trauma patients admitted from 2008 to 2010 who sustained no abdominal pain or abnormalities on physical examination of the abdomen at admission and were submitted to computed tomography of the abdomen and/or exploratory laparotomy. Patients were assigned into: Group 1 (with intra-abdominal injuries) or Group 2 (without intra-abdominal injuries). Variables were compared between groups to identify those significantly associated with the presence of intra-abdominal injuries, adopting p<0.05 as significant. Subsequently, the variables with p<0.20 on bivariate analysis were selected to create a logistic regression model using the forward stepwise method. Results: A total of 268 cases met the inclusion criteria. Patients in Group I were characterized as having significantly (p<0.05) lower mean AIS score for the head segment (1.0±1.4 vs. 1.8±1.9), as well as higher mean AIS thorax score (1.6±1.7 vs. 0.9±1.5) and ISS (25.7±14.5 vs. 17,1±13,1). The rate of abdominal injuries was significantly higher in run-over pedestrians (37.3%) and in motorcyclists (36.0%) (p<0.001). The resultant logistic regression model provided 73.5% accuracy for identifying abdominal injuries. The variables included were: motorcyclist accident as trauma mechanism (p<0.001 - OR 5.51; 95%CI 2.40-12.64), presence of rib fractures (p<0.003 - OR 3.00; 95%CI 1.47-6.14), run-over pedestrian as trauma mechanism (p=0.008 - OR 2.85; 95%CI 1.13-6.22) and abnormal neurological physical exam at admission (p=0.015 - OR 0.44; 95%CI 0.22-0.85). Conclusion Intra-abdominal injuries were predominantly associated with trauma mechanism and presence of chest injuries.

Diarrhea outbreaks in suckling piglets due to rotavirus group C single and mixed (rotavirus groups A and B) infections

Porcine group A rotavirus (PoRVA) is a major cause of neonatal diarrhea in suckling and recently weaned piglets worldwide. The involvement of non-group A rotavirus in cases of neonatal diarrhea in piglets are sporadic. In Brazil there are no reports of the porcine rotavirus group C (PoRVC) as etiologic agent of the diarrhea outbreaks in piglets. The aim of this study was to describe the identification of rotavirus group C in single and in mixed infection with rotavirus groups A and B in three neonatal diarrhea outbreaks in suckling (<21-day-old) piglets, with 70% to 80% and 20% to 25% of morbidity and lethality rates, respectively, in three pig herds located in the state of Santa Catarina, Brazil. The diagnosis of PoRV in the diarrheic fecal samples was performed using polyacrylamide gel electrophoresis (PAGE) to identify the presence of porcine rotavirus groups A, B (PoRVB), and C, and by RT-PCR (PoRVA and PoRVC) and semi-nested (SN)-PCR (PoRVB) to partially amplify the VP4 (VP8*)-VP7, NSP2, and VP6 genes of PoRVA, PoRVB, and PoRVC, respectively. One RT-PCR (PoRVA and PoRVC) and SN-PCR (PoRVB) product of each group of rotavirus of each diarrhea outbreak was submitted to nucleotide (nt) sequence analysis. Based on the PAGE technique, 4 (25%) and 1 (6.25%) of the 16 diarrheic fecal samples evaluated in the first outbreak presented PoRVA and PoRVC electropherotype, respectively, and 11 (68.75%) were negative. In the second outbreak, 3 (42.85%) of the 7 fecal samples evaluated presented PoRVA electropherotype, and in 3 (42.85%) and in 1 (14.3%) fecal samples were detected inconclusive and negative results, respectively. Three (30%) of the 10 fecal samples of the third outbreak presented PoRVC electropherotype; 5 (50%) and 2 (20%) samples showed negative and inconclusive results, respectively. Based on the RT-PCR and SN-PCR assays in the first neonatal diarrhea outbreak, PoRVC was detected in 13 (81.2%) of the 16 diarrheic fecal samples evaluated. PoRVC single infection was identified in 4 (25%) of these samples and mixed infections with PoRVA and PoRVB in 9 (56.2%) fecal samples. All of the seven diarrheic fecal samples evaluated from the second neonatal diarrhea outbreak were positive for PoRVC, whereas its mixed infection with other PoRV groups was detected in 4 (57.2%) samples. In the third outbreak, PoRVC in single infection was detected in all of the 10 diarrheic fecal samples analyzed. In the nt sequence analysis, the PoRVA strains of the first and second outbreaks demonstrated higher nt identity with G4P[6] and G9P[23] genotypes, respectively. The PoRVB strains (first and second outbreaks) and the PoRVC strains (first, second, and third outbreaks) showed higher nt identity and clustered in the phylogenetic tree with PoRVB and PoRVC strains that belong to the N4 and I1 genotypes, respectively. This is the first description in Brazil of the involvement of PoRVC in the etiology of diarrhea outbreaks in suckling piglets. The results of this study demonstrated that PoRVC, in both single and mixed infections, is an important enteropathogen involved in neonatal diarrhea outbreaks in piglets and that the use of more sensitive diagnostic techniques allows the identification of mixed infections involving two or even three groups of PoRV, which may be more common than previously reported.

Switchable Lanthanide Luminescence for Detection of Biomolecules

Binary probes are oligonucleotide probe pairs that hybridize adjacently to a complementary target nucleic acid. In order to detect this hybridization, the two probes can be modified with, for example, fluorescent molecules, chemically reactive groups or nucleic acid enzymes. The benefit of this kind of binary probe based approach is that the hybridization elicits a detectable signal which is distinguishable from background noise even though unbound probes are not removed by washing before measurement. In addition, the requirement of two simultaneous binding events increases specificity. Similarly to binary oligonucleotide probes, also certain enzymes and fluorescent proteins can be divided into two parts and used in separation-free assays. Split enzyme and fluorescent protein reporters have practical applications among others as tools to investigate protein-protein interactions within living cells. In this study, a novel label technology, switchable lanthanide luminescence, was introduced and used successfully in model assays for nucleic acid and protein detection. This label technology is based on a luminescent lanthanide chelate divided into two inherently non-luminescent moieties, an ion carrier chelate and a light harvesting antenna ligand. These form a highly luminescent complex when brought into close proximity; i.e., the label moieties switch from a dark state to a luminescent state. This kind of mixed lanthanide complex has the same beneficial photophysical properties as the more typical lanthanide chelates and cryptates - sharp emission peaks, long emission lifetime enabling time-resolved measurement, and large Stokes’ shift, which minimize the background signal. Furthermore, the switchable lanthanide luminescence technique enables a homogeneous assay set-up. Here, switchable lanthanide luminescence label technology was first applied to sensitive, homogeneous, single-target nucleic acid and protein assays with picomolar detection limits and high signal to background ratios. Thereafter, a homogeneous four-plex nucleic acid array-based assay was developed. Finally, the label technology was shown to be effective in discrimination of single nucleotide mismatched targets from fully matched targets and the luminescent complex formation was analyzed more thoroughly. In conclusion, this study demonstrates that the switchable lanthanide luminescencebased label technology can be used in various homogeneous bioanalytical assays.

Membrane development for oil contaminated water treatments

The aim of this Master’s thesis study was to develop a membrane for oil contaminated water treatments. Oily wastewaters are a big problem to environment and therefore it is important to find an efficient method for their treatment. There are several treatment methods, but one of the most promising methods is membrane filtration. In the theoretical part of this study the membrane technology and polymeric membrane preparation with phase inversion and membrane modification methods was discussed. It was also told about the most important properties of the membranes. Oily waters, their treatment methods and oily wastewater sources were discussed more specifically. In the experimental part membranes from cellulose acetate were prepared and membranes were modified with two different methods. Modification methods were surface modification and polymer mixing. The modification purpose was to make membranes more hydrophilic and increase surface charge, which can reduce fouling. Membranes were characterized by determining zeta potential, contact angle, oil retention, pure water permeability, pressure-normalized flux and fouling. It were used both synthetic and real spent oil-water emulsion in membrane filtration. Surface modification resulted membranes, which had better properties than unmodified membrane. The amount of substance used in surface modification affected a lot to membrane properties, so it would be necessary to try different amounts of substance to develop the best membrane for oil-water emulsion treatment.

Inheritance of anthracnose resistance in common bean genotypes P.I. 207262 and AB 136

Bean (Phaseolus vulgaris) lines P.I. 207262 and AB 136, both resistant to delta and kappa races of Colletotrichum lindemuthianum, were crossed with Michelite, Dark Red Kidney, and Perry Marrow, susceptible to both races, and with Cornell 49-242, resistant to delta and susceptible to kappa. F1 and F2 reactions demonstrated that P.I. 207262 carries duplicate dominant genes for resistance to the delta race; AB 136 carries a dominant gene. These resistance genes are independent of the Are gene from Cornell 49-242. With respect to the kappa race, F1 and F2 data showed that the resistance controlled by P.I. 207262 and by AB 136 depends on a single dominant gene. Complementary factors were involved with AB 136 resistance to the delta race and with P.I. 207262 resistance to kappa.

Environmental color affects Nile tilapia reproduction

We investigated the effects of environmental color on the reproductive behavior of Nile tilapia, Oreochromis niloticus. Two environmental colors were tested by covering the aquarium (60 x 60 x 40 cm) with white (12 groups) or blue (13 groups) cellophane and observing reproductive behavior in groups of 2 males (10.27 ± 0.45 cm) and 3 females (10.78 ± 0.45 cm) each. After assignment to the respective environmental color (similar luminosity = 100 to 120 Lux), the animals were observed until reproduction (identified by eggs in the female's mouth) or up to 10 days after the first nest building. Photoperiod was from 6:00 h to 18:00 h every day. Food was offered in excess once a day and water quality was similar among aquaria. Daily observations were made at 8:00, 11:00, 14:00 and 17:00 h regarding: a) latency to the first nest, b) number of nests, c) gravel weight removed (the male excavates the nest in the bottom of the aquarium), d) nest area, and e) mouthbrooding incubation (indication of reproduction). The proportion of reproducing fish was significantly higher (6 of 13) in the group exposed to the blue color compared the group exposed to the white color (1 of 12; Goodman's test of proportions). Moreover, males under blue light removed significantly larger masses of gravel (blue = 310.70 ± 343.50 g > white = 130.38 ± 102.70 g; P = 0.01) and constructed wider nests (blue = 207.93 ± 207.80 cm² > white = 97.68 ± 70.64 cm²; P = 0.03) than the control (white). The other parameters did not differ significantly between light conditions. We concluded that reproduction in the presence of blue light was more frequent and intense than in the presence of white light.

Efficacy of HUMN criteria for scoring the micronucleus assay in human lymphocytes exposed to a low concentration of p,p'-DDT

The cytokinesis-block micronucleus (CBMN) assay is one of the standard cytogenetic tools employed to assess chromosomal damage subsequent to exposure to genotoxic/cytotoxic agents, and is widely applicable to plant, animal and human cells. In the present study, the CBMN assay was used to assess the baseline damage in binuclear human peripheral blood lymphocytes exposed to 25 µg/L p,p'-DDT for 1, 2, 24, and 48 h by measuring the frequency of micronuclei, nucleoplasmic bridges and nuclear buds. These new scoring criteria facilitated the detection of different types of clastogenic and aneugenic effects induced by this type of pollutant. With these criteria, CBMN can also be used to measure nucleoplasmic bridges which are considered to be consequences of chromosome rearrangements and nuclear buds which are biomarkers of altered gene amplification and gene dosage. The total number of micronuclei observed in binuclear human peripheral blood lymphocytes of the exposed samples (ranging from 32 to 47) was significantly greater (P < 0.05) than that detected in the unexposed (0 time) control sample, where the total number of micronuclei was 7. The number of nucleoplasmic bridges and nuclear buds obtained after 24 and 48 h was also significantly (P < 0.05) greater in the samples treated with p,p'-DDT than in the unexposed control samples. Thus, our results confirmed the usefulness of the new criteria applicable for the CBMN assay employed in measuring the DNA damage and its role of a sensitive cytogenetic biomarker.

p16INK4 expression in precursor lesions of squamous cell cervical cancer related to the presence of HPV-DNA

The purpose of the present study was to identify the expression of p16INK4 in cervical cancer precursor lesions by immunohistochemistry and to correlate it with lesion grade and presence of human papillomavirus (HPV) infection. Cervical specimens from 144 women seen consecutively at the gynecology outpatient clinic of our institution from December 2003 to May 2005 were analyzed by cytopathology, histopathology, polymerase chain reaction for HPV-DNA, and p16INK4 immunostaining. Histologically normal biopsies, HPV-DNA negative by polymerase chain reaction, were used as control. HPV-DNA prevalence, including the control group, was 68.1% and the prevalence of p16INK4 expression was 55.0%. The percentage of cells stained by p16INK4 ranged from 10 to 100%, both in the group consisting of cervical intraepithelial neoplasia (CIN)1/HPV specimens and in the group of CIN2/CIN3 specimens with P value of 0.0001. p16INK4 expression was 48.3% in the CIN1/HPV group, as opposed to 94.3% in the CIN2/CIN3 group (P = 0.001), showing a statistically significant difference between the two groups. The quantitative method used here is simple and less subjective than the different semiquantitative methods described in the literature. In view of the different definitions of a p16INK4-positive case, it is almost impossible to compare the findings reported by different investigators. This study confirms the association between p16INK4 and CIN2 and CIN3 lesions. Moreover, it shows that some low grade lesions expressed high levels of this protein. This may indicate that such low grade lesions may be predisposed to progress to high grade lesions. This means that p16INK4 may be a strong marker for "neoplastic lesions" induced by HPV and not just an infection marker.

Increased levels of glutamate in the central nervous system are associated with behavioral symptoms in experimental malaria

Cerebral malaria (CM) is a severe complication resulting from Plasmodium falciparum infection. This condition has been associated with cognitive, behavioral and motor dysfunctions, seizures and coma. The underlying mechanisms of CM are incompletely understood. Glutamate and other metabolites such as lactate have been implicated in its pathogenesis. In the present study, we investigated the involvement of glutamate in the behavioral symptoms of CM. Seventeen female C57BL/6 mice (20-25 g) aged 6-8 weeks were infected with P. berghei ANKA by the intraperitoneal route using a standardized inoculation of 10(6) parasitized red blood cells suspended in 0.2 mL PBS. Control animals (N = 17) received the same volume of PBS. Behavioral and neurological symptoms were analyzed by the SmithKline/Harwell/Imperial College/Royal Hospital/Phenotype Assessment (SHIRPA) battery. Glutamate release was measured in the cerebral cortex and cerebrospinal fluid of infected and control mice by fluorimetric assay. All functional categories of the SHIRPA battery were significantly altered in the infected mice at 6 days post-infection (dpi) (P ≤ 0.05). In parallel to CM symptoms, we found a significant increase in glutamate levels in the cerebral cortex (mean ± SEM; control: 11.62 ± 0.90 nmol/mg protein; infected at 3 dpi: 10.36 ± 1.17 nmol/mg protein; infected at 6 dpi: 26.65 ± 0.73 nmol/mg protein; with EGTA, control: 5.60 ± 1.92 nmol/mg protein; infected at 3 dpi: 6.24 ± 1.87 nmol/mg protein; infected at 6 dpi: 14.14 ± 0.84 nmol/mg protein) and in the cerebrospinal fluid (control: 128 ± 51.23 pmol/mg protein; infected: 301.4 ± 22.52 pmol/mg protein) of infected mice (P ≤ 0.05). These findings suggest a role of glutamate in the central nervous system dysfunction found in CM.

Relationship between genetic mutation variations and acute-phase reactants in the attack-free period of children diagnosed with familial Mediterranean fever

Familial Mediterranean fever (FMF) is a periodic autoinflammatory disease characterized by chronic inflammation. This study investigated the relationship between acute-phase reactants and gene mutations in attack-free periods of childhood FMF. Patients diagnosed with FMF were divided into four groups based on genetic features: no mutation, homozygous, heterozygous, and compound heterozygous. These groups were monitored for 2 years, and blood samples were collected every 6 months during attack-free periods. Erythrocyte sedimentation rate, C-reactive protein, fibrinogen, and white blood cell count were measured. A disease severity score was determined for each patient. Mean values for erythrocyte sedimentation rate and fibrinogen were significantly different in the homozygous group. White blood cell count and C-reactive protein were similar between the groups. Disease severity score was higher in patients with the M694V mutation than in individuals without the mutation, as well as in those with other mutation groups. Periodic follow-up of patients with FMF MEFV mutations in subjects with acute-phase reactants may be useful in the prevention of morbidity.

Immobilization and therapeutic passive stretching generate thickening and increase the expression of laminin and dystrophin in skeletal muscle

Extracellular matrix and costamere proteins transmit the concentric, isometric, and eccentric forces produced by active muscle contraction. The expression of these proteins after application of passive tension stimuli to muscle remains unknown. This study investigated the expression of laminin and dystrophin in the soleus muscle of rats immobilized with the right ankle in plantar flexion for 10 days and subsequent remobilization, either by isolated free movement in a cage or associated with passive stretching for up to 10 days. The intensity of the macrophage response was also evaluated. One hundred and twenty-eight female Wistar rats were divided into 8 groups: free for 10 days; immobilized for 10 days; immobilized/free for 1, 3, or 10 days; or immobilized/stretched/free for 1, 3, or 10 days. After the experimental procedures, muscle tissue was processed for immunofluorescence (dystrophin/laminin/CD68) and Western blot analysis (dystrophin/laminin). Immobilization increased the expression of dystrophin and laminin but did not alter the number of macrophages in the muscle. In the stretched muscle groups, there was an increase in dystrophin and the number of macrophages after 3 days compared with the other groups; dystrophin showed a discontinuous labeling pattern, and laminin was found in the intracellular space. The amount of laminin was increased in the muscles treated by immobilization followed by free movement for 10 days. In the initial stages of postimmobilization (1 and 3 days), an exacerbated macrophage response and an increase of dystrophin suggested that the therapeutic stretching technique induced additional stress in the muscle fibers and costameres.

Socially shared metacognitive regulation during collaborative learning processes in student dyads and small groups

Traditionally metacognition has been theorised, methodologically studied and empirically tested from the standpoint mainly of individuals and their learning contexts. In this dissertation the emergence of metacognition is analysed more broadly. The aim of the dissertation was to explore socially shared metacognitive regulation (SSMR) as part of collaborative learning processes taking place in student dyads and small learning groups. The specific aims were to extend the concept of individual metacognition to SSMR, to develop methods to capture and analyse SSMR and to validate the usefulness of the concept of SSMR in two different learning contexts; in face-to-face student dyads solving mathematical word problems and also in small groups taking part in inquiry-based science learning in an asynchronous computer-supported collaborative learning (CSCL) environment. This dissertation is comprised of four studies. In Study I, the main aim was to explore if and how metacognition emerges during problem solving in student dyads and then to develop a method for analysing the social level of awareness, monitoring, and regulatory processes emerging during the problem solving. Two dyads comprised of 10-year-old students who were high-achieving especially in mathematical word problem solving and reading comprehension were involved in the study. An in-depth case analysis was conducted. Data consisted of over 16 (30–45 minutes) videotaped and transcribed face-to-face sessions. The dyads solved altogether 151 mathematical word problems of different difficulty levels in a game-format learning environment. The interaction flowchart was used in the analysis to uncover socially shared metacognition. Interviews (also stimulated recall interviews) were conducted in order to obtain further information about socially shared metacognition. The findings showed the emergence of metacognition in a collaborative learning context in a way that cannot solely be explained by individual conception. The concept of socially-shared metacognition (SSMR) was proposed. The results highlighted the emergence of socially shared metacognition specifically in problems where dyads encountered challenges. Small verbal and nonverbal signals between students also triggered the emergence of socially shared metacognition. Additionally, one dyad implemented a system whereby they shared metacognitive regulation based on their strengths in learning. Overall, the findings suggested that in order to discover patterns of socially shared metacognition, it is important to investigate metacognition over time. However, it was concluded that more research on socially shared metacognition, from larger data sets, is needed. These findings formed the basis of the second study. In Study II, the specific aim was to investigate whether socially shared metacognition can be reliably identified from a large dataset of collaborative face-to-face mathematical word problem solving sessions by student dyads. We specifically examined different difficulty levels of tasks as well as the function and focus of socially shared metacognition. Furthermore, the presence of observable metacognitive experiences at the beginning of socially shared metacognition was explored. Four dyads participated in the study. Each dyad was comprised of high-achieving 10-year-old students, ranked in the top 11% of their fourth grade peers (n=393). Dyads were from the same data set as in Study I. The dyads worked face-to-face in a computer-supported, game-format learning environment. Problem-solving processes for 251 tasks at three difficulty levels taking place during 56 (30–45 minutes) lessons were video-taped and analysed. Baseline data for this study were 14 675 turns of transcribed verbal and nonverbal behaviours observed in four study dyads. The micro-level analysis illustrated how participants moved between different channels of communication (individual and interpersonal). The unit of analysis was a set of turns, referred to as an ‘episode’. The results indicated that socially shared metacognition and its function and focus, as well as the appearance of metacognitive experiences can be defined in a reliable way from a larger data set by independent coders. A comparison of the different difficulty levels of the problems suggested that in order to trigger socially shared metacognition in small groups, the problems should be more difficult, as opposed to moderately difficult or easy. Although socially shared metacognition was found in collaborative face-to-face problem solving among high-achieving student dyads, more research is needed in different contexts. This consideration created the basis of the research on socially shared metacognition in Studies III and IV. In Study III, the aim was to expand the research on SSMR from face-to-face mathematical problem solving in student dyads to inquiry-based science learning among small groups in an asynchronous computer-supported collaborative learning (CSCL) environment. The specific aims were to investigate SSMR’s evolvement and functions in a CSCL environment and to explore how SSMR emerges at different phases of the inquiry process. Finally, individual student participation in SSMR during the process was studied. An in-depth explanatory case study of one small group of four girls aged 12 years was carried out. The girls attended a class that has an entrance examination and conducts a language-enriched curriculum. The small group solved complex science problems in an asynchronous CSCL environment, participating in research-like processes of inquiry during 22 lessons (á 45–minute). Students’ network discussion were recorded in written notes (N=640) which were used as study data. A set of notes, referred to here as a ‘thread’, was used as the unit of analysis. The inter-coder agreement was regarded as substantial. The results indicated that SSMR emerges in a small group’s asynchronous CSCL inquiry process in the science domain. Hence, the results of Study III were in line with the previous Study I and Study II and revealed that metacognition cannot be reduced to the individual level alone. The findings also confirm that SSMR should be examined as a process, since SSMR can evolve during different phases and that different SSMR threads overlapped and intertwined. Although the classification of SSMR’s functions was applicable in the context of CSCL in a small group, the dominant function was different in the asynchronous CSCL inquiry in the small group in a science activity than in mathematical word problem solving among student dyads (Study II). Further, the use of different analytical methods provided complementary findings about students’ participation in SSMR. The findings suggest that it is not enough to code just a single written note or simply to examine who has the largest number of notes in the SSMR thread but also to examine the connections between the notes. As the findings of the present study are based on an in-depth analysis of a single small group, further cases were examined in Study IV, as well as looking at the SSMR’s focus, which was also studied in a face-to-face context. In Study IV, the general aim was to investigate the emergence of SSMR with a larger data set from an asynchronous CSCL inquiry process in small student groups carrying out science activities. The specific aims were to study the emergence of SSMR in the different phases of the process, students’ participation in SSMR, and the relation of SSMR’s focus to the quality of outcomes, which was not explored in previous studies. The participants were 12-year-old students from the same class as in Study III. Five small groups consisting of four students and one of five students (N=25) were involved in the study. The small groups solved ill-defined science problems in an asynchronous CSCL environment, participating in research-like processes of inquiry over a total period of 22 hours. Written notes (N=4088) detailed the network discussions of the small groups and these constituted the study data. With these notes, SSMR threads were explored. As in Study III, the thread was used as the unit of analysis. In total, 332 notes were classified as forming 41 SSMR threads. Inter-coder agreement was assessed by three coders in the different phases of the analysis and found to be reliable. Multiple methods of analysis were used. Results showed that SSMR emerged in all the asynchronous CSCL inquiry processes in the small groups. However, the findings did not reveal any significantly changing trend in the emergence of SSMR during the process. As a main trend, the number of notes included in SSMR threads differed significantly in different phases of the process and small groups differed from each other. Although student participation was seen as highly dispersed between the students, there were differences between students and small groups. Furthermore, the findings indicated that the amount of SSMR during the process or participation structure did not explain the differences in the quality of outcomes for the groups. Rather, when SSMRs were focused on understanding and procedural matters, it was associated with achieving high quality learning outcomes. In turn, when SSMRs were focused on incidental and procedural matters, it was associated with low level learning outcomes. Hence, the findings imply that the focus of any emerging SSMR is crucial to the quality of the learning outcomes. Moreover, the findings encourage the use of multiple research methods for studying SSMR. In total, the four studies convincingly indicate that a phenomenon of socially shared metacognitive regulation also exists. This means that it was possible to define the concept of SSMR theoretically, to investigate it methodologically and to validate it empirically in two different learning contexts across dyads and small groups. In-depth micro-level case analysis in Studies I and III showed the possibility to capture and analyse in detail SSMR during the collaborative process, while in Studies II and IV, the analysis validated the emergence of SSMR in larger data sets. Hence, validation was tested both between two environments and within the same environments with further cases. As a part of this dissertation, SSMR’s detailed functions and foci were revealed. Moreover, the findings showed the important role of observable metacognitive experiences as the starting point of SSMRs. It was apparent that problems dealt with by the groups should be rather difficult if SSMR is to be made clearly visible. Further, individual students’ participation was found to differ between students and groups. The multiple research methods employed revealed supplementary findings regarding SSMR. Finally, when SSMR was focused on understanding and procedural matters, this was seen to lead to higher quality learning outcomes. Socially shared metacognition regulation should therefore be taken into consideration in students’ collaborative learning at school similarly to how an individual’s metacognition is taken into account in individual learning.