Purification of pharmaceuticals and nutraceutical compounds by sub- and supercritical extraction and chromatography

This thesis discusses the use of sub- and supercritical fluids as the medium in extraction and chromatography. Super- and subcritical extraction was used to separate essential oils from herbal plant Angelica archangelica. The effect of extraction parameters was studied and sensory analyses of the extracts were done by an expert panel. The results of the sensory analyses were compared to the analytically determined contents of the extracts. Sub- and supercritical fluid chromatography (SFC) was used to separate and purify high-value pharmaceuticals. Chiral SFC was used to separate the enantiomers of racemic mixtures of pharmaceutical compounds. Very low (cryogenic) temperatures were applied to substantially enhance the separation efficiency of chiral SFC. The thermodynamic aspects affecting the resolving ability of chiral stationary phases are briefly reviewed. The process production rate which is a key factor in industrial chromatography was optimized by empirical multivariate methods. General linear model was used to optimize the separation of omega-3 fatty acid ethyl esters from esterized fish oil by using reversed-phase SFC. Chiral separation of racemic mixtures of guaifenesin and ferulic acid dimer ethyl ester was optimized by using response surface method with three variables per time. It was found that by optimizing four variables (temperature, load, flowate and modifier content) the production rate of the chiral resolution of racemic guaifenesin by cryogenic SFC could be increased severalfold compared to published results of similar application. A novel pressure-compensated design of industrial high pressure chromatographic column was introduced, using the technology developed in building the deep-sea submersibles (Mir 1 and 2). A demonstration SFC plant was built and the immunosuppressant drug cyclosporine A was purified to meet the requirements of US Pharmacopoeia. A smaller semi-pilot size column with similar design was used for cryogenic chiral separation of aromatase inhibitor Finrozole for use in its development phase 2.

Sensory auditory processing and intuitive sound detection : an investigation of musical experts and nonexperts

The auditory system can detect occasional changes (deviants) in acoustic regularities without the need for subjects to focus their attention on the sound material. Deviant detection is reflected in the elicitation of the mismatch negativity component (MMN) of the event-related potentials. In the studies presented in this thesis, the MMN is used to investigate the auditory abilities for detecting similarities and regularities in sound streams. To investigate the limits of these processes, professional musicians have been tested in some of the studies. The results show that auditory grouping is already more advanced in musicians than in nonmusicians and that the auditory system of musicians can, unlike that of nonmusicians, detect a numerical regularity of always four tones in a series. These results suggest that sensory auditory processing in musicians is not only a fine tuning of universal abilities, but is also qualitatively more advanced than in nonmusicians. In addition, the relationship between the auditory change-detection function and perception is examined. It is shown that, contrary to the generally accepted view, MMN elicitation does not necessarily correlate with perception. The outcome of the auditory change-detection function can be implicit and the implicit knowledge of the sound structure can, after training, be utilized for behaviorally correct intuitive sound detection. These results illustrate the automatic character of the sensory change detection function.

Neuronal oscillations in gamma- and alpha-frequency bands: from object representations to sensory awareness

The synchronization of neuronal activity, especially in the beta- (14-30 Hz) /gamma- (30 80 Hz) frequency bands, is thought to provide a means for the integration of anatomically distributed processing and for the formation of transient neuronal assemblies. Thus non-stimulus locked (i.e. induced) gamma-band oscillations are believed to underlie feature binding and the formation of neuronal object representations. On the other hand, the functional roles of neuronal oscillations in slower theta- (4 8 Hz) and alpha- (8 14 Hz) frequency bands remain controversial. In addition, early stimulus-locked activity has been largely ignored, as it is believed to reflect merely the physical properties of sensory stimuli. With human neuromagnetic recordings, both the functional roles of gamma- and alpha-band oscillations and the significance of early stimulus-locked activity in neuronal processing were examined in this thesis. Study I of this thesis shows that even the stimulus-locked (evoked) gamma oscillations were sensitive to high-level stimulus features for speech and non-speech sounds, suggesting that they may underlie the formation of early neuronal object representations for stimuli with a behavioural relevance. Study II shows that neuronal processing for consciously perceived and unperceived stimuli differed as early as 30 ms after stimulus onset. This study also showed that the alpha band oscillations selectively correlated with conscious perception. Study III, in turn, shows that prestimulus alpha-band oscillations influence the subsequent detection and processing of sensory stimuli. Further, in Study IV, we asked whether phase synchronization between distinct frequency bands is present in cortical circuits. This study revealed prominent task-sensitive phase synchrony between alpha and beta/gamma oscillations. Finally, the implications of Studies II, III, and IV to the broader scientific context are analysed in the last study of this thesis (V). I suggest, in this thesis that neuronal processing may be extremely fast and that the evoked response is important for cognitive processes. I also propose that alpha oscillations define the global neuronal workspace of perception, action, and consciousness and, further, that cross-frequency synchronization is required for the integration of neuronal object representations into global neuronal workspace.

Crystallization as a Tool for Controlling and Designing Properties of Pharmaceutical Solids

The ability to deliver the drug to the patient in a safe, efficacious and cost-effective manner depends largely on the physicochemical properties of the active pharmaceutical ingredient (API) in the solid state. In this context, crystallization is of critical importance in pharmaceutical industry, as it defines physical and powder properties of crystalline APIs. An improved knowledge of the various aspects of crystallization process is therefore needed. The overall goal of this thesis was to gain better understanding of the relationships between crystallization, solid-state form and properties of pharmaceutical solids with a focus on a crystal engineering approach to design technological properties of APIs. Specifically, solid-state properties of the crystalline forms of the model APIs, erythromycin A and baclofen, and the influence of solvent on their crystallization behavior were investigated. In addition, the physical phenomena associated with wet granulation and hot-melting processing of the model APIs were examined at the molecular level. Finally, the effect of crystal habit modification of a model API on its tabletting properties was evaluated. The thesis enabled the understanding of the relationship between the crystalline forms of the model APIs, which is of practical importance for solid-state control during processing and storage. Moreover, a new crystalline form, baclofen monohydrate, was discovered and characterized. Upon polymorph screening, erythromycin A demonstrated high solvate-forming propensity thus emphasizing the need for careful control of the solvent effects during formulation. The solvent compositions that yield the desirable crystalline form of erythromycin A were defined. Furthermore, new examples on solvent-mediated phase transformations taking place during wet granulation of baclofen and hot-melt processing of erythromycin A dihydrate with PEG 6000 are reported. Since solvent-mediated phase transformations involve the crystallization of a stable phase and hence affect the dissolution kinetics and possibly absorption of the API these transformations must be well documented. Finally, a controlled-crystallization method utilizing HPMC as a crystal habit modifier was developed for erythromycin A dihydrate. The crystals with modified habit were shown to posses improved compaction properties as compared with those of unmodified crystals. This result supports the idea of morphological crystal engineering as a tool for designing technological properties of APIs and is of utmost practical interest.

Constructive Perspectives on Inductive Logic

Constructive (intuitionist, anti-realist) semantics has thus far been lacking an adequate concept of truth in infinity concerning factual (i.e., empirical, non-mathematical) sentences. One consequence of this problem is the difficulty of incorporating inductive reasoning in constructive semantics. It is not possible to formulate a notion for probable truth in infinity if there is no adequate notion of what truth in infinity is. One needs a notion of a constructive possible world based on sensory experience. Moreover, a constructive probability measure must be defined over these constructively possible empirical worlds. This study defines a particular kind of approach to the concept of truth in infinity for Rudolf Carnap's inductive logic. The new approach is based on truth in the consecutive finite domains of individuals. This concept will be given a constructive interpretation. What can be verifiably said about an empirical statement with respect to this concept of truth, will be explained, for which purpose a constructive notion of epistemic probability will be introduced. The aim of this study is also to improve Carnap's inductive logic. The study addresses the problem of justifying the use of an "inductivist" method in Carnap's lambda-continuum. A correction rule for adjusting the inductive method itself in the course of obtaining evidence will be introduced. Together with the constructive interpretation of probability, the correction rule yields positive prior probabilities for universal generalizations in infinite domains.

Cortical processing of speech and non-speech sounds in adults and newborns

Comprehension of a complex acoustic signal - speech - is vital for human communication, with numerous brain processes required to convert the acoustics into an intelligible message. In four studies in the present thesis, cortical correlates for different stages of speech processing in a mature linguistic system of adults were investigated. In two further studies, developmental aspects of cortical specialisation and its plasticity in adults were examined. In the present studies, electroencephalographic (EEG) and magnetoencephalographic (MEG) recordings of the mismatch negativity (MMN) response elicited by changes in repetitive unattended auditory events and the phonological mismatch negativity (PMN) response elicited by unexpected speech sounds in attended speech inputs served as the main indicators of cortical processes. Changes in speech sounds elicited the MMNm, the magnetic equivalent of the electric MMN, that differed in generator loci and strength from those elicited by comparable changes in non-speech sounds, suggesting intra- and interhemispheric specialisation in the processing of speech and non-speech sounds at an early automatic processing level. This neuronal specialisation for the mother tongue was also reflected in the more efficient formation of stimulus representations in auditory sensory memory for typical native-language speech sounds compared with those formed for unfamiliar, non-prototype speech sounds and simple tones. Further, adding a speech or non-speech sound context to syllable changes was found to modulate the MMNm strength differently in the left and right hemispheres. Following the acoustic-phonetic processing of speech input, phonological effort related to the selection of possible lexical (word) candidates was linked with distinct left-hemisphere neuronal populations. In summary, the results suggest functional specialisation in the neuronal substrates underlying different levels of speech processing. Subsequently, plasticity of the brain's mature linguistic system was investigated in adults, in whom representations for an aurally-mediated communication system, Morse code, were found to develop within the same hemisphere where representations for the native-language speech sounds were already located. Finally, recording and localization of the MMNm response to changes in speech sounds was successfully accomplished in newborn infants, encouraging future MEG investigations on, for example, the state of neuronal specialisation at birth.

Cortical processing of musical pitch as reflected by behavioural and electrophysiological evidence

In a musical context, the pitch of sounds is encoded according to domain-general principles not confined to music or even to audition overall but common to other perceptual and cognitive processes (such as multiple pattern encoding and feature integration), and to domain-specific and culture-specific properties related to a particular musical system only (such as the pitch steps of the Western tonal system). The studies included in this thesis shed light on the processing stages during which pitch encoding occurs on the basis of both domain-general and music-specific properties, and elucidate the putative brain mechanisms underlying pitch-related music perception. Study I showed, in subjects without formal musical education, that the pitch and timbre of multiple sounds are integrated as unified object representations in sensory memory before attentional intervention. Similarly, multiple pattern pitches are simultaneously maintained in non-musicians' sensory memory (Study II). These findings demonstrate the degree of sophistication of pitch processing at the sensory memory stage, requiring neither attention nor any special expertise of the subjects. Furthermore, music- and culture-specific properties, such as the pitch steps of the equal-tempered musical scale, are automatically discriminated in sensory memory even by subjects without formal musical education (Studies III and IV). The cognitive processing of pitch according to culture-specific musical-scale schemata hence occurs as early as at the sensory-memory stage of pitch analysis. Exposure and cortical plasticity seem to be involved in musical pitch encoding. For instance, after only one hour of laboratory training, the neural representations of pitch in the auditory cortex are altered (Study V). However, faulty brain mechanisms for attentive processing of fine-grained pitch steps lead to inborn deficits in music perception and recognition such as those encountered in congenital amusia (Study VI). These findings suggest that predispositions for exact pitch-step discrimination together with long-term exposure to music govern the acquisition of the automatized schematic knowledge of the music of a particular culture that even non-musicians possess.

Processing of Stimulus Repetition and Change in the Somatosensory System: Recordings of Electrical and Magnetic Brain Responses

In the present work, effects of stimulus repetition and change in a continuous stimulus stream on the processing of somatosensory information in the human brain were studied. Human scalp-recorded somatosensory event-related potentials (ERPs) and magnetoencephalographic (MEG) responses rapidly diminished with stimulus repetition when mechanical or electric stimuli were applied to fingers. On the contrary, when the ERPs and multi-unit a ctivity (MUA) were directly recorded from the primary (SI) and secondary (SII) somatosensory cortices in a monkey, there was no marked decrement in the somatosensory responses as a function of stimulus repetition. These results suggest that this rate effect is not due to the response diminution in the SI and SII cortices. Obviously the responses to the first stimulus after a long "silent" period are nhanced due to unspecific initial orientation, originating in more broadly distributed and/or deeper neural structures, perhaps in the prefrontal cortices. With fast repetition rates not only the late unspecific but also some early specific somatosensory ERPs were diminished in amplitude. The fast decrease of the ERPs as a function of stimulus repetition is mainly due to the disappearance of the orientation effect and with faster repetition rates additively due to stimulus specific refractoriness. A sudden infrequent change in the continuous stimulus stream also enhanced somatosensory MEG responses to electric stimuli applied to different fingers. These responses were quite similar to those elicited by the deviant stimuli alone when the frequent standard stimuli were omitted. This enhancement was obviously due to the release from refractoriness because the neural structures generating the responses to the infrequent deviants had more time to recover from the refractoriness than the respective structures for the standards. Infrequent deviant mechanical stimuli among frequent standard stimuli also enhanced somatosensory ERPs and, in addition, they elicited a new negative wave which did not occur in the deviants-alone condition. This extra negativity could be recorded to deviations in the stimulation site and in the frequency of the vibratory stimuli. This response is probably a somatosensory analogue of the auditory mismatch negativity (MMN) which has been suggested to reflect a neural mismatch process between the sensory input and the sensory memory trace.

Minimalistinen yhdyssanateoria ja käsitteiden yhdistely

The thesis aims to link the biolinguistic research program and the results of studies in comceptual combination from cognitive psychology. The thesis derives a theory of syntactic structure of noun and adjectival compounds from the Empty Lexicon Hypothesis. Two compound-forming operations are described: root-compounding and word-compounding. The aptness of theory is tested with finnish and greek compounds. From the syntactic theory semantic requirements for conceptual system are derived, especially requirements for handling morphosyntactic features. These requirements are compared to three formidable theories of conceptual combination: relation theory CARIN, Dual-Process theory and C3-theory. The claims of explanatory power of relational distributions of modifier in CARIN-theory ared discarded, as the method for sampling and building relational distributions is not reliable and the algorithmic instantiation of theory does not compute what it claims to compute. From relational theory there still remains results supporting existence of 'easy' relations for certain concepts. Dual-Process theory is found to provide results that cannot in theory be affected by linguistic system, but the basic idea of property compounds is kept. C3-theory is found to be not computationally realistic, but the basic results of diagnosticity and local properties (domains) of conceptual system are solid. The three conceptual combination models are rethought as a problem of finding the shortest route between the two concepts. The new basis for modeling is suggested to be bare conceptual landscape with morphosyntactiic or semantic features working as guidance and structural features of landscape basically unknown, but such as they react to features from linguistic system. Minimalistic principles to conceptual modeling are suggested.

Haptiisit ja hapteemit : tapaustutkimus kuurosokean henkilön kosketukseen perustuvan kommunikaation kehityksestä

Haptices and haptemes: A case study of developmental process in touch-based communication of acquired deafblind people This research is the first systematic, longitudinal process and development description of communication using touch and body with an acquired deafblind person. The research consists of observational and analysed written and video materials mainly from two informants´ experiences during period of 14 years. The research describes the adaptation of Social-Haptic methods between a couple, and other informants´ experiences, which have been collated from biographies and through giving national and international courses. When the hearing and sight deteriorates due to having an acquired deafblind condition, communication consists of multi-systematic and adaptive methods. A person`s expressive language, spoken or Sign Language, usually remains unchanged, but the methods of receiving information could change many times during a person s lifetime. Haptices are made from haptemes that determines which regulations are analysed. When defining haptemes the definition, classification and varied meanings of touch were discovered. Haptices include sharing a personal body space, meaning of touch-contact, context and using different communication channels. Communication distances are classified as exact distance, estimated distance and touch distance. Physical distance can be termed as very long, long, medium or very close. Social body space includes the body areas involved in sending and receiving haptices and applying different types of contacts. One or two hands can produce messages by using different hand shapes and orientations. This research classifies how the body can be identified into different areas such as body orientation, varied body postures, body position levels, social actions and which side of the body is used. Spatial body space includes environmental and situational elements. Haptemes of movements are recognised as the direction of movements, change of directions on the body, directions between people, pressure, speed, frequency, size, length, duration, pause, change of rhythm, shape, macro and micro movements. Haptices share multidimensional meanings and emotions. Research describes haptices in different situations enhancing sensory information and functioning also as an independent language. Haptices includes social-haptic confirmation system, social quick messages, body drawing, contact to the people and the environment, guiding and sharing art experiences through movements. Five stages of emotional differentiation were identified as very light, light, medium, heavy and very heavy touch. Haptices give the possibility to share different art, hobby and game experiences. A new communication system development based on the analysis of the research data is classified into different phases. These are experimental initiation, social deconstruction, developing the description of Social-Haptic communication and generalisation of the theory as well as finding and conceptualising the haptices and haptemes. The use and description of haptices is a social innovation, which illustrates the adaptive function of the body and perceptual senses that can be taught to a third party. Keywords: deafblindness, hapteme, haptic, haptices, movement, social-haptic communication, social-haptic confirmation system, tactile, touch

Lukemisen vaikeuden kuntoutus ensiluokkalaisilla : Kolme pedagogista interventiota

Remediation of Reading Difficulties in Grade 1. Three Pedagogical Interventions Keywords: initial teaching, learning to read, reading difficulties, intervention, dyslexia, remediation of dyslexia, home reading, computerized training In this study three different reading interventions were tested for first-graders at risk of reading difficulties at school commencement. The intervention groups were compared together and with a control group receiving special education provided by the school. First intervention was a new approach called syllable rhythmics in which syllabic rhythm, phonological knowledge and letter-phoneme correspondence are emphasized. Syllable rhythmics is based on multi-sensory training elements aimed at finding the most functional modality for every child. The second intervention was computerized training of letter-sound correspondence with the Ekapeli learning game. The third intervention was home-based shared book reading, where every family was given a story book, and dialogic reading style reading and writing exercises were prepared for each chapter of the book. The participants were 80 first-graders in 19 classes in nine schools. The children were matched in four groups according to pre-test results: three intervention and one control. The interventions took ten weeks starting from September in grade 1. The first post-test including several measures of reading abilities was administered in December. The first delayed post-test was administered in March, the second in September in grade 2, and the third, “ALLU” test (reading test for primary school) was administered in March in grade 2. The intervention and control groups differed only slightly from each other in grade 1. However, girls progressed significantly more than boys in both word reading and reading comprehension in December and this difference remained in March. The children who had been cited as inattentive by their teachers also lagged behind the others in the post-tests in December and March. When participants were divided into two groups according to their initial letter knowledge at school entry, the weaker group (maximum 17 correctly named letters in pre-test) progressed more slowly in both word reading and reading comprehension in grade 1. Intervention group and gender had no interaction effect in grade 1. Instead, intervention group and attentiveness had an interaction effect on most test measures the inattentive students in the syllable rhythmic group doing worst and attentive students in the control group doing best in grade 1. The smallest difference between results of attentive and inattentive students was in the Ekapeli group. In grade 2 still only minor differences were found between the intervention groups and control group. The only significant difference was in non-word reading, with the syllable rhythmics group outperforming the other groups in the fall. The difference between girls’ and boys’ performances in both technical reading and text comprehension disappeared in grade 2. The difference between the inattentive and attentive students cold no longer be found in technical reading, and the difference became smaller in text comprehension as well. The difference between two groups divided according to their initial letter knowledge disappeared in technical reading but remained significant in text comprehension measures in the ALLU test in the spring of grade 2. In all, the children in the study did better in the ALLU test than expected according to ALLU test norms. Being the weakest readers in their classes in the pre-test, 52.3 % reached the normal reading ability level. In the norm group 72.3 % of all students attained normal reading ability. The results of this study indicate that different types of remediation programs can be effective, and that special education has been apparently useful. The results suggest careful consideration of first-graders’ initial reading abilities (especially letter knowledge) and possible failure of attention; remediation should be individually targeted while flexibly using different methods.

Kuva tikkeinä kankaalla : Kokemuksia PE-design 6.0 -kirjontakuvion suunnitteluohjelman kahden automaattisen tekniikan käytettävyydestä

The aim of the thesis was to compare the correspondence of the outcome a computer assisted program appearance compared to the original image. The aspect of the study was directed to embroidery with household machines. The study was made from the usability point of view with Brother's PE-design 6.0 embroidery design programs two automatic techniques; multicoloured fragment design and multicoloured stitch surface design. The study's subject is very current because of the fast development of machine embroidery. The theory is based on history of household sewing machines, embroidery sewing machines, stitch types in household sewing machines, embroidery design programs as well as PE-design 6.0 embroidery design program's six automatic techniques. Additionally designing of embroidery designs were included: original image, digitizing, punching, applicable sewing threads as well as the connection between embroidery designs and materials used on embroidery. Correspondences of sewn appearances were examined with sewing experimental methods. 18 research samples of five original image were sewn with both techniques. Experiments were divided into four testing stages in design program. Every testing stage was followed by experimental sewing with Brother Super Galaxie 3100D embroidery machine. Experiments were reported into process files and forms made for the techniques. Research samples were analysed on images syntactic bases with sensory perception assessment. Original images and correspondence of the embroidery appearances were analysed with a form made of it. The form was divided into colour and shape assessment in five stage-similarity-scale. Based on this correspondence analysis it can be said that with both automatic techniques the best correspondence of colour and shape was achieved by changing the standard settings and using the makers own thread chart and edited original image. According to the testing made it is impossible to inform where the image editing possibilities of the images are sufficient or does the optimum correspondence need a separate program. When aiming at correspondence between appearances of two images the computer is unable to trace by itself the appearance of the original image. Processing a computer program assisted embroidery image human perception and personal decision making are unavoidable.