Towards desired crystalline product properties: In-situ monitoring of batch crystallization

The objective of industrial crystallization is to obtain a crystalline product which has the desired crystal size distribution, mean crystal size, crystal shape, purity, polymorphic and pseudopolymorphic form. Effective control of the product quality requires an understanding of the thermodynamics of the crystallizing system and the effects of operation parameters on the crystalline product properties. Therefore, obtaining reliable in-line information about crystal properties and supersaturation, which is the driving force of crystallization, would be very advantageous. Advanced techniques, such asRaman spectroscopy, attenuated total reflection Fourier transform infrared (ATR FTIR) spectroscopy, and in-line imaging techniques, offer great potential for obtaining reliable information during crystallization, and thus giving a better understanding of the fundamental mechanisms (nucleation and crystal growth) involved. In the present work, the relative stability of anhydrate and dihydrate carbamazepine in mixed solvents containing water and ethanol were investigated. The kinetics of the solvent mediated phase transformation of the anhydrate to hydrate in the mixed solvents was studied using an in-line Raman immersion probe. The effects of the operation parameters in terms of solvent composition, temperature and the use of certain additives on the phase transformation kineticswere explored. Comparison of the off-line measured solute concentration and the solid-phase composition measured by in-line Raman spectroscopy allowedthe identification of the fundamental processes during the phase transformation. The effects of thermodynamic and kinetic factors on the anhydrate/hydrate phase of carbamazepine crystals during cooling crystallization were also investigated. The effect of certain additives on the batch cooling crystallization of potassium dihydrogen phosphate (KDP) wasinvestigated. The crystal growth rate of a certain crystal face was determined from images taken with an in-line video microscope. An in-line image processing method was developed to characterize the size and shape of thecrystals. An ATR FTIR and a laser reflection particle size analyzer were used to study the effects of cooling modes and seeding parameters onthe final crystal size distribution of an organic compound C15. Based on the obtained results, an operation condition was proposed which gives improved product property in terms of increased mean crystal size and narrowersize distribution.

In situ spectroellipsometric study of the nucleation and growth of amorphous silicon

A detailed in situ spectroellipsometric analysis of the nucleation and growth of hydrogenated amorphous silicon (a:Si:H) is presented. Photoelectronic quality a‐Si:H films are deposited by plasma‐enhanced chemical vapor deposition on smooth metal (NiCr alloy) and crystalline silicon (c‐Si) substrates. The deposition of a‐Si:H is analyzed from the first monolayer up to a final thickness of 1.2 μm. In order to perform an improved analysis, real time ellipsometric trajectories are recorded, using fixed preparation conditions, at various photon energies ranging from 2.2 to 3.6 eV. The advantage of using such a spectroscopic experimental procedure is underlined. New insights into the nucleation and growth mechanisms of a‐Si:H are obtained. The nucleation mechanism on metal and c‐Si substrates is very accurately described assuming a columnar microstructural development during the early stage of the growth. Then, as a consequence of the incomplete coalescence of the initial nuclei, a surface roughness at the 10-15 Å scale is identified during the further growth of a‐Si:H on both substrates. The bulk a‐Si:H grows homogeneously beneath the surface roughness. Finally, an increase of the surface roughness is evidenced during the long term growth of a‐Si:H. However, the nature of the substrate influenced the film growth. In particular, the film thickness involved in the nucleation‐coalescence phase is found lower in the case of c‐Si (67±8 Å) as compared to NiCr (118±22 Å). Likewise films deposited on c‐Si present a smaller surface roughness even if thick samples are considered (>1 μm). More generally, the present study illustrates the capability of in situ spectroellipsometry to precisely analyze fundamental processes in thin‐film growth, but also to monitor the preparation of complex structures on a few monolayers scale.

Técnicas in situ de baixo custo em eletroquímica: a microbalança a cristal de quartzo

Among in situ techniques, the electrochemical quartz crystal microbalance (EQCM) is a powerful tool for the study of electrochemical reactions that produce mass changes in the electrode/solution interface. This review present some systems in which the EQCM combined with classical electrochemical techniques, gives relevant information for understanding the charge transport process at a molecular level. The aim of this review is to do a brief description of experimental arrangements, with emphasis on some special cares that must be considered by the users. Secondly, some chosen electrochemical systems where the technique was successfully applied are discussed. Finally, a brief analysis of electroacoustic impedance experiments was done in order to show when the Sauerbrey equation can be used.

Rapid increase in incidence of breast ductal cancer in situ in Girona, Spain 1983-2007

Introduction: The aim of this study was to describe breast ductal cancer in situ (DCIS) incidence trends in women in the Girona province during a period of 25 years. The influence of age, use of mammography and implementation of the breast cancer screening programs was explored. Incidence of subsequent invasive breast cancers (IBC) and DCIS treatment was also considered. Materials and Methods: Cases diagnosed with primary pure DCIS (n=416) during 1983-2007 were extracted from the population-based Girona Cancer Registry. The estimated annual percent change was estimated using joinpoint analysis. Results: DCIS incidence showed a sharp rise until 1998, followed by a less marked upward trend. Among women aged 50-69 the increase was particularly important between 1992 and in 1996, reflecting the spread in mammography use. Conclusion: The upward trend of DCIS was mainly related to an increase in mammography use either opportunistic or as a result of screening implementation

Comparison of nuclear grade and immunohistochemical features in situ and invasive components of ductal carcinoma of breast

PURPOSE:To compare the prognostic and predictive features between in situ and invasive components of ductal breast carcinomas. METHODS:We selected 146 consecutive breast samples with ductal carcinoma in situ (DCIS) associated with adjacent invasive breast carcinoma (IBC). We evaluated nuclear grade and immunohistochemical expression of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), cytokeratin 5/6 (CK5/6), and epidermal growth factor receptor (EGFR) in both components, in situ and invasive, and the Ki-67 percentage of cells in the invasive part. The DCIS and IBC were classified in molecular surrogate types determined by the immunohistochemical profile as luminal (RE/PR-positive/ HER2-negative), triple-positive (RE/RP/HER2-positive), HER2-enriched (ER/PR-negative/HER2-positive), and triple-negative (RE/RP/HER2-negative). Discrimination between luminal A and luminal B was not performed due to statistical purposes. Correlations between the categories in the two groups were made using the Spearman correlation method. RESULTS:There was a significant correlation between nuclear grade (p<0.0001), expression of RE/RP (p<0.0001), overexpression of HER2 (p<0.0001), expression of EGFR (p<0.0001), and molecular profile (p<0.0001) between components in situ and IBC. CK 5/6 showed different distribution in DCIS and IBC, presenting a significant association with the triple-negative phenotype in IBC, but a negative association among DCIS. CONCLUSIONS: Our results suggest that classical prognostic and predictive features of IBC are already determined in the preinvasive stage of the disease. However the role of CK5/6 in invasive carcinoma may be different from the precursor lesions.

Acute promyelocytic leukemia: the study of t(15;17) translocation by fluorescent in situ hybridization, reverse transcriptase-polymerase chain reaction and cytogenetic techniques

Acute promyelocytic leukemia (AML M3) is a well-defined subtype of leukemia with specific and peculiar characteristics. Immediate identification of t(15;17) or the PML/RARA gene rearrangement is fundamental for treatment. The objective of the present study was to compare fluorescent in situ hybridization (FISH), reverse transcriptase-polymerase chain reaction (RT-PCR) and karyotyping in 18 samples (12 at diagnosis and 6 after treatment) from 13 AML M3 patients. Bone marrow samples were submitted to karyotype G-banding, FISH and RT-PCR. At diagnosis, cytogenetics was successful in 10 of 12 samples, 8 with t(15;17) and 2 without. FISH was positive in 11/12 cases (one had no cells for analysis) and positivity varied from 25 to 93% (mean: 56%). RT-PCR was done in 6/12 cases and all were positive. Four of 8 patients with t(15;17) presented positive RT-PCR as well as 2 without metaphases. The lack of RT-PCR results in the other samples was due to poor quality RNA. When the three tests were compared at diagnosis, karyotyping presented the translocation in 80% of the tested samples while FISH and RT-PCR showed the PML/RARA rearrangement in 100% of them. Of 6 samples evaluated after treatment, 3 showed a normal karyotype, 1 persistence of an abnormal clone and 2 no metaphases. FISH was negative in 4 samples studied and 2 had no material for analysis. RT-PCR was positive in 4 (2 of which showed negative FISH, indicating residual disease) and negative in 2. When the three tests were compared after treatment, they showed concordance in 2 of 6 samples or, when there were not enough cells for all tests, concordance between karyotype and RT-PCR in one. At remission, RT-PCR was the most sensitive test in detecting residual disease, as expected (positive in 4/6 samples). An incidence of about 40% of 5' breaks and 60% of 3' breaks, i.e., bcr3 and bcr1/bcr2, respectively, was observed.

Localization of monoterpenoid indole alkaloid (MIA) biosynthesis by in situ hybridization (ISH)

Monoterpenoid indole alkaloids (MIA) are among the largest and most complex group of nitrogen containing secondary metabolites that are characteristic of the Apocynaceae plant family including the most notable Catharanthus roseus. These compounds have demonstrated activity as successful drugs for treating various cancers, neurological disorders and cardiovascular conditions. Due to the low yields of these compounds and high pharmacological value, their biosynthesis is a major topic of study. Previous work highlighting the leaf epidermis and leaf surface as a highly active area in MIA biosynthesis and MIA accumulation has made the epidermis a major focus of this thesis. This thesis provides an in-depth analysis of the valuable technique of RNA in situ hybridization (ISH) and demonstrates the application of the technique to analyze the location of the biosynthetic steps involved in the production of MIAs. The work presented in this thesis demonstrates that most of the MIAs of Eurasian Vinca minor, African Tabernaemontana e/egans and five Amsonia species, including North American Amsonia hubrichitii and Mediterranean A. orienta/is, accumulate in leaf wax exudates, while the rest of the leaf is almost devoid of alkaloids. Biochemical studies on Vinca minor displayed high tryptophan decarboxylase (TOe) enzyme activity and protein expression in the leaf epidermis compared to whole leaves. ISH studies aimed at localizing TOe and strictosidine synthase suggest the upper and lower epidermis of V. minor and T. e/egans as probable significant production sites for MIAs that will accumulate on the leaf surface, however the results don't eliminate the possibility of the involvement of other cell types. The monoterpenoid precursor to all MIAs, secologanin, is produced through the MEP pathway occurring in two cell types, the IPAP cells (Gl0H) and epidermal cells (LAMT and SLS). The work presented in this thesis, localizes a novel enzymatic step, UDPG-7-deoxyloganetic acid glucosyltransferase (UGT8) to the IPAP cells of Catharanthus longifolius. These results enable the suggestion that all steps from Gl0H up to and including UGT8 occur in the IPAP cells of the leaf, making the IPAP cells the main site for the majority of secologanin biosynthesis. It also makes the IPAP cells a likely cell type to begin searching for the gene of the uncharacterized steps between Gl0H and UGT8. It also narrows the compound to be transported from the IPAP cells to either 7-deoxyloganic acid or loganic acid, which aids in the identification of the transportation mechanism.

Tratamiento del deslizamiento epifisiario capital femoral con fijación in situ versus luxación quirúrgica

Introducción: El Deslizamiento Epifisiario Capital Femoral es la enfermedad de la cadera más común en adolescentes entre los 9 y 16 años. Es de causa idiopática, más frecuente en hombres, se clasifica en 4 estadios según criterios clínicos y radiológicos. Se buscó evaluar la evolución de los deslizamientos moderados y severos tratados con una de las dos técnicas propuestas. Metodología Se realizó un estudio descriptivo con pacientes que fueron llevados a fijación in situ o luxación controlada entre 2008 y 2011. Resultados: Se incluyeron 26 pacientes, los cuales el 65.4% se les realizó luxación quirúrgica controlada y el 34.6% fijación in situ. El 70,6% de pacientes tenían DECF inestable y 70,5% tenían desplazamiento severo. La evaluación de la escala WOMAC para dolor, rigidez y capacidad funcional encontró mejores beneficios para el grupo de fijación in situ, estadísticamente significativos (p<0,05), no solo en términos de dolor, rigidez y capacidad funcional sino menor frecuencia de complicaciones. Las complicaciones más frecuentes en el grupo de luxación quirúrgica controlada fueron un caso de infección, 7 casos (41,2%) de necrosis avascular de cabeza femoral, 5 casos (29,4%) de condrolisis y 2 casos (11,8%) de pseudoartrosis; En el grupo de fijación in situ, solo 1 (11,1%) presentó Infección del Sitio Operatorio y 1 (11,1%) Condrolisis. Resultados significativos solo para necrosis avascular. Discusión: Los pacientes con deslizamientos moderados y severos manejados con fijación in situ tuvieron una mejor resultado con menor proporción de complicaciones.

Intermittent release of transients in the slow solar wind: 2. In situ evidence

In paper 1, we showed that the Heliospheric Imager (HI) instruments on the pair of NASA STEREO spacecraft can be used to image the streamer belt and, in particular, the variability of the slow solar wind which originates near helmet streamers. The observation of intense intermittent transient outflow by HI implies that the corresponding in situ observations of the slow solar wind and corotating interaction regions (CIRs) should contain many signatures of transients. In the present paper, we compare the HI observations with in situ measurements from the STEREO and ACE spacecraft. Analysis of the solar wind ion, magnetic field, and suprathermal electron flux measurements from the STEREO spacecraft reveals the presence of both closed and partially disconnected interplanetary magnetic field lines permeating the slow solar wind. We predict that one of the transients embedded within the second CIR (CIR‐D in paper 1) should impact the near‐Earth ACE spacecraft. ACE measurements confirm the presence of a transient at the time of CIR passage; the transient signature includes helical magnetic fields and bidirectional suprathermal electrons. On the same day, a strahl electron dropout is observed at STEREO‐B, correlated with the passage of a high plasma beta structure. Unlike ACE, STEREO‐B observes the transient a few hours ahead of the CIR. STEREO‐A, STEREO‐B, and ACE spacecraft observe very different slow solar wind properties ahead of and during the CIR analyzed in this paper, which we associate with the intermittent release of transients.

Incorporation of in situ and biomarker assays in higher-tier assessment of the aquatic toxicity of insecticides

This study was designed to test the feasibility of integrating in situ, single species exposures and biomarker analysis into microcosm studies. Experimental ponds were dosed with pirimiphos methyl (PM) and lindane. C. riparius fourth instar larvae were deployed for 48 h on nine separate occasions during the study period before and after treatment. Surviving larvae were analysed for acetylcholinesterase activity (AChE). Survival and biomarker data were compared to chironomid assemblage analysis by monitoring insects emerging from the microcosms. Survival of chironomids within the in situ systems commenced on day + 16 after treatment with 31.6% and 53.3% survival in the lindane and PM treated ponds, respectively. In contrast, the first emergence from the microcosms occurred on days + 27, in respect to lindane, and + 59 for the PM treated ponds. Thus the in situ bioassay was able to demonstrate gradual reduction in toxicity within the sediment before this was evident from macroinvertebrate monitoring. Significant ACNE inhibition was only detected on exposure to PM. Levels decreased from 75% on day + 16 to 26% by day +29. The biomarker analysis confirmed that, by the end of the study, the insecticide was no longer exerting an effect. We discuss how the use of in situ bioassays could also aid comparison of microcosm studies by adding a standardized dimension. (C) 2003 Elsevier Ltd. All rights reserved.

Complete filling of zeolite frameworks with polyalkynes formed in situ by transition-metal ion catalysts

We report the use of transition-metal-exchanged zeolites as media for the catalytic formation and encapsulation of both polyethyne and polypropyne, and computer modeling studies on the composites so formed. Alkyne gas was absorbed into the pores of zeolite Y (Faujasite) exchanged with transition-metal cations [Fe(II), Co(II), Cu(II), Ni(II), and Zn(II)]. Ni(II) and Zn(II) were found to be the most efficient for the production of poly-ynes. These cations were also found to be effective in polymer generation when exchanged in zeolites mordenite and beta. The resulting powdered samples were characterized by FTIR, Raman, diffuse reflectance electronic spectroscopy, TEM, and elemental analysis, revealing, nearly complete loading of the zeolite channels for the majority of the samples. Based on the experimental carbon content, we have derived the percentage of channel filling, and the proportion of the channels containing a single polymer chain for mordenite. Experimentally, the channels for Y are close to complete filling for polyethyne (PE) and polypropyne (PP), and this is also true for polyethyne in mordenite. Computer modeling studies using Cerius2 show that the channels of mordenite can only accept a single polymer chain of PP, in which case these channels are also completely filled.

Structure of single-wall peptide nanotubes: in situ flow aligning X-ray diffraction

The structure of single wall peptide nanotubes is presented for the model surfactant-like peptide A6K. Capillary flow alignment of a sample in the nematic phase at high concentration in water leads to oriented X-ray diffraction patterns. Analysis of these, accompanied by molecular dynamics simulations, suggests the favourable self-assembly of antiparallel peptide dimers into beta-sheet ribbons that wrap helically to form the nanotube wall.

Longitudinal investigation of the faecal microbiota of healthy full-term infants using fluorescence in situ hybridization and denaturing gradient gel electrophoresis.

From birth onwards, the gastrointestinal (GI) tract of infants progressively acquires a complex range of micro-organisms. It is thought that by 2 years of age the GI microbial population has stabilized. Within the developmental period of the infant GI microbiota, weaning is considered to be most critical, as the infant switches from a milk-based diet (breast and/or formula) to a variety of food components. Longitudinal analysis of the biological succession of the infant GI/faecal microbiota is lacking. In this study, faecal samples were obtained regularly from 14 infants from 1 month to 18 months of age. Seven of the infants (including a set of twins) were exclusively breast-fed and seven were exclusively formula-fed prior to weaning, with 175 and 154 faecal samples, respectively, obtained from each group. Diversity and dynamics of the infant faecal microbiota were analysed by using fluorescence in situ hybridization and denaturing gradient gel electrophoresis. Overall, the data demonstrated large inter- and intra-individual differences in the faecal microbiological profiles during the study period. However, the infant faecal microbiota merged with time towards a climax community within and between feeding groups. Data from the twins showed the highest degree of similarity both quantitatively and qualitatively. Inter-individual variation was evident within the infant faecal microbiota and its development, even within exclusively formula-fed infants receiving the same diet. These data can be of help to future clinical trials (e.g. targeted weaning products) to organize protocols and obtain a more accurate outline of the changes and dynamics of the infant GI microbiota.

Representativity of in situ precipitation measurements - a case study for the LITFASS area in North-Eastern Germany

In situ precipitation measurements can extremely differ in space and time. Taking into account the limited spatial–temporal representativity and the uncertainty of a single station is important for validating mesoscale numerical model results as well as for interpreting remote sensing data. In situ precipitation data from a high resolution network in North-Eastern Germany are analysed to determine their temporal and spatial representativity. For the dry year 2003 precipitation amounts were available with 10 min resolution from 14 rain gauges distributed in an area of 25 km 25 km around the Meteorological Observatory Lindenberg (Richard-Aßmann Observatory). Our analysis reveals that short-term (up to 6 h) precipitation events dominate (94% of all events) and that the distribution is skewed with a high frequency of very low precipitation amounts. Long-lasting precipitation events are rare (6% of all precipitation events), but account for nearly 50% of the annual precipitation. The spatial representativity of a single-site measurement increases slightly for longer measurement intervals and the variability decreases. Hourly precipitation amounts are representative for an area of 11 km 11 km. Daily precipitation amounts appear to be reliable with an uncertainty factor of 3.3 for an area of 25 km 25 km, and weekly and monthly precipitation amounts have uncertainties of a factor of 2 and 1.4 when compared to 25 km 25 km mean values.

In situ signatures of interchange reconnection between magnetic clouds and open magnetic fields: a mechanism for the erosion of polar coronal holes?

We outline a method to determine the direction of solar open flux transport that results from the opening of magnetic clouds (MCs) by interchange reconnection at the Sun based solely on in-situ observations. This method uses established findings about i) the locations and magnetic polarities of emerging MC footpoints, ii) the hemispheric dependence of the helicity of MCs, and iii) the occurrence of interchange reconnection at the Sun being signaled by uni-directional suprathermal electrons inside MCs. Combining those observational facts in a statistical analysis of MCs during solar cycle 23 (period 1995 – 2007), we show that the time of disappearance of the northern polar coronal hole (1998 – 1999), permeated by an outward-pointing magnetic field, is associated with a peak in the number of MCs originating from the northern hemisphere and connected to the Sun by outward-pointing magnetic field lines. A similar peak is observed in the number of MCs originating from the southern hemisphere and connected to the Sun by inward-pointing magnetic field lines. This pattern is interpreted as the result of interchange reconnection occurring between MCs and the open field lines of nearby polar coronal holes. This reconnection process closes down polar coronal hole open field lines and transports these open field lines equatorward, thus contributing to the global coronal magnetic field reversal process. These results will be further constrainable with the rising phase of solar cycle 24.