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We recently published the 3-month follow-up of 2 neonates with Robin sequence whose mandibular hypoplasia and restricted airway were successfully treated with an orthodontic airway plate (OAP) without surgical intervention. Both infants were successfully weaned off the OAP after several months of continuous use. We present the course of OAP treatment in these patients with a focus on breathing, feeding, and facial growth during their first year of life. Both infants demonstrated stable mandibular projection, resolution of obstructive sleep apnea, and normal development.

Various Achilleaspecies are rich in bioactive compounds and are important medicinal plants in phytotherapy. In the present study, Achillea millefoliumL., Achillea moschataWulfen, and Achillea atrataL. were compared with respect to their phenolic profile and antibacterial activity against gram-positive bacteria strains (Staphylococcus, Propionibacterium). Particular focus was given to A. atrata, which has hardly been studied so far. Based on the metabolite profile, A. atrataexhibited more similarities to A. moschatathan to A. millefolium. The former two only differed in the occurrence of four compounds. The flavonols syringetin-3-O-glucoside and mearnsetin-hexoside, not reported for an Achilleaspecies before, have been detected in A. atrataand A. moschata. All Achilleaspecies reduced growth of the tested bacteria. A. atratademonstrated highest activity against Propionibacterium acnesand Staphylococcus epidermidis, both being involved in the pathogenesis of acne vulgaris. Furthermore, A. atratahas a pronounced anti–methicillin-resistant Staphylococcus aureuspotential. Bioassay-guided fractionation revealed that only the most polar fraction of A. moschatadisplayed antimicrobial activity, which was attributed to phenolics such as apigenin, centaureidin, and nevadensin, being present in high amounts in A. atrata. Thus, this alpine species shows promising antimicrobial activity and might be a potential source for developing novel dermal/topical drugs.

Essential oils are widely used in the food and cosmetics industry as natural flavoring and fragrance substances. For this reason, a thorough quality control applying selected analytical methods is required. Oxidation along with hydroperoxide formation is an important drawback during production and storage of essential oils. Hydroperoxides constitute the main products formed upon photo-oxidation of essential oils. Due to hydroperoxide instability, gas chromatography (GC) and high-performance liquid chromatography (HPLC) analyses are required. According to the European Pharmacopoeia, titration is the official method for oxidation assessment. However, this analysis is time-consuming, and large sample quantities are required. Here, we present a simple and accurate spectrophotometric method for the detection of peroxide trace amounts in essential oils and terpenes. The principle is based on the formation of Wurster’s red, which is enforced by the peroxide-driven oxidation of N,N-dimethyl-p-phenylenediamine dihydrochloride (DMPD). The method was validated using dibenzoyl peroxide (DBP) and cumene hydroperoxide (CHP). To demonstrate the suitability of the method for routine analysis, various oxidized terpenes and essential oils were chosen. Moreover, photo- and thermal oxidation experiments were compared and evaluated using gas chromatography/mass spectrometry (GC/MS) and a synthesized limonene-2-hydroperoxide (Lim-2-OOH) reference standard to gather detailed information on the structural changes of the respective terpenes.

Intrathecal immunoglobulin G (IgG) synthesis and oligoclonal IgG bands in cerebrospinal fluid (CSF) are hallmarks of multiple sclerosis (MS), but the antigen specificities remain enigmatic. Our study is the first investigating the autoantibody repertoire in paired serum and CSF samples from patients with relapsing-remitting MS (RRMS), primary progressive MS (PPMS), and other neurological diseases by the use of high-density peptide microarrays. Protein sequences of 45 presumed MS autoantigens (e.g.MOG, MBP, and MAG) were represented on the microarrays by overlapping 15mer peptides. IgG reactivities were screened against a total of 3991 peptides, including also selected viral epitopes. The measured antibody reactivities were highly individual but correlated for matched serum and CSF samples. We found 54 peptides to be recognized significantly more often by serum or CSF antibodies from MS patients compared with controls (pvalues <0.05). The results for RRMS and PPMS clearly overlapped. However, PPMS patients presented a broader peptide-antibody signature. The highest signals were detected for a peptide mapping to a region of the Epstein-Barr virus protein EBNA1 (amino acids 392–411), which is homologous to the N-terminal part of human crystallin alpha-B. Our data confirmed several known MS-associated antigens and epitopes, and they delivered additional potential linear epitopes, which await further validation. The peripheral and intrathecal humoral immune response in MS is polyspecific and includes antibodies that are also found in serum of patients with other diseases. Further studies are required to assess the pathogenic relevance of autoreactive and anti-EBNA1 antibodies as well as their combinatorial value as biomarkers for MS.

The performance of a passive air surveillance sensor based on Global Navigation Satellite Systems (GNSS) is mainly limited by the receiver noise efficiency, the achievable signal processing gain and the radar cross section (RCS) of an airplane. For surveillance applications large detection ranges as well as a high probability of detection are crucial parameters. Due to the very low GNSS signal powers received on the earth’s surface, high radar cross sections are mandatory to achieve detection ranges for airplanes at some kilometers distance. This paper will discuss a multistatic transmitter and receiver arrangement, which is indispensable to get a reasonable detection rate with respect to a hemispheric field of view. The strong performance dependency of such a sensor on the number of transmitters and receivers will be shown by means of some exemplary simulation results.

Wireless transmission systems in smart metering networks share the advantage of lower installation costs due to the expandability of separate infrastructure but suffer from transmission problems. In this paper the issue of interference of wireless transmitted smart meter data with third party systems and data from other meters is investigated and an approach for solving the problem is presented. A multi-channel wireless m-bus receiver was developed to separate the desired data from unwanted interferers by spatial filtering. The according algorithms are presented and the influence of different antenna types on the spatial filtering is investigated. The performance of the spatial filtering is evaluated by extensive measurements in a realistic surrounding with several hundreds of active wireless m-bus transponders. These measurements correspond to the future environment for data-collectors as they took place in rural and urban areas with smart gas meters equipped with wireless m-bus transponders installed in almost all surrounding buildings.

A solidified ionic liquid matrix (SILM) consisting of 3-aminoquinoline, a-cyano-4-hydroxycinnamic acid and ammonium dihydrogen phosphate combines the benefits of liquid and solid MALDI matrices and proves to be well suitable for phosphopeptide analysis using MALDI-MS in the low femtomole range. Desalting and buffer exchange that typically follow after phosphopeptide elution from metal oxide affinity chromatography (MOAC) materials can be omitted. Shifting the pH from acidic to basic during target preparation causes slow matrix crystallization and homogeneous embedding of the analyte molecules, forming a uniform preparation from which (phospho) peptides can be ionized in high yields over long periods of time. The novel combination of MOAC-based phosphopeptide enrichment with SILM preparation has been developed with commercially available standard phosphopeptides and with a-casein as phosphorylated standard protein. The applicability of the streamlined phosphopeptide analysis procedure to cell biological and clinical samples has been tested (i) using affinity-enriched endogenous TRIM28 from cell cultures and (ii) by analysis of a two-dimensional gel-separated protein spot from a bladder cancer sample.

Patent Intelligence analysis provides an overview of the patent activity in the technical fields and giving insights into the competitors` pipeline and is a starting point for preparing own product, intellectual property strategies and development efforts. The Patent Intelligence analysis help to discover new application, serving catalyst a new thinking and creative approaches to business problem or identifying strategic growth opportunities.

This paper presents a novel non-destructive method to locate and size damages in frame structures, performed by examining and interpreting changes in measured vibration response. The method bases on a relation, prior contrived by the authors, between the strain energy distribution in the structure for the transversal vibration modes and the modal changes (in terms of natural frequencies) due to damage. Using this relation a damage location indicator DLI was derived, which permits to locate cracks in spatial structures. In this paper an L-frame is considered for proving the applicability of this method. First the mathematical expressions for the modes shapes and their derivatives were determined and simulation result compared with that obtained by finite element analysis. Afterwards patterns characterizing damage locations were derived and compared with measurement results on the real structure; the DLI permitted accurate localization of any crack placed in the two structural elements.

There have been many successful efforts to enrich phosphopeptides in complex protein mixtures by the use of immobilized metal affinity chromatography (IMAC) and/or metal oxide affinity chromatography (MOAC) with which mass spectrometric analysis of phosphopeptides has become state of the art in specialized laboratories, mostly applying nanoLC electrospray ionization mass spectrometry-based investigations. However, widespread use of these powerful techniques is still not achieved. In this study, we present a ready-to-use phosphopeptide enrichment procedure using commercially available TiO2-loaded pipette tips in combination with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analyses. Using β-casein as a model protein and citric acid as additive during sample loading, a similar enrichment success can be achieved as compared to applying 2,5-dihydroxy benzoic acid (DHB) for this task. But the DHB-inherited drawbacks are eliminated. In addition, we show that combining DHB and 2,4,6-trihydroxy acetophenone (THAP) as matrix for MALDI-MS measurements retains the sensitivity of DHB for phosphopeptide analysis but adds the homogenous crystallization properties of THAP, enabling preparation of evenly distributed matrix surfaces on MALDI-MS anchor targets, a prerequisite for automated MALDI-MS analyses. Tripartite motif-containing protein 28 and stathmin are two examples for which successful phosphopeptide enrichment of either sodium dodecyl sulfate polyacrylamide gel electrophoresis or two-dimensional gel electrophoresis-separated proteins is shown. Finally, high resolution MALDI Fourier transform ion cyclotron resonance mass spectrometry after phosphopeptide enrichment suggests that chemical dephosphorylation may occur as a side reaction during basic elution of phosphopeptides bound to MOAC surfaces, suggesting that proteome-wide phosphopeptide analyses ought to be interpreted with caution. In contrast, in-depth analysis of phosphopeptide/non-phosphorylated peptide siblings may be used to estimate stability differences of phosphorylation sites in individual proteins, possibly adding valuable information on biological regulation processes.

Investigation of the dichloromethane extracts from herbal and root parts of Mercurialis perennis L. afforded a mixture of 11 homologous n-alkylresorcinols (ARs) with saturated odd-numbered alkyl side chains (C15:0-C27:0). In addition to three predominant ARs (C19:0, C21:0 and C23:0), a number of minor ARs were identified by use of LC-MS/MS and GC-MS techniques. Among the compounds detected, four uncommon ARs with evennumbered alkyl side chain lengths were also determined. The overall AR concentration in herbal parts was 7 to 9 times higher compared to that of the roots. The results presented may open a new view on the phytochemistry and pharmacognosy of M. perennis and other members of the Euphorbiaceae family.

Two new scandium platinum silicides, ScPt2Si (orthorhombic, Cmcm, a= 3.6653(6) Å, b= 9.831(1) Å, c= 6.967(1) Å, Z= 4) and Sc5Pt9Si7(hexagonal, P$\bar{6}$, a= 9.9418(5) Å, c= 3.8832(2) Å, Z= 1) were prepared by reaction of the elements in an electric arc furnace and subsequent annealing. The crystal structures were solved and refined from single crystal diffraction data. ScPt2Si crystallizes with an ‘inverse’ MgCuAl2structure whereas Sc5Pt9Si7forms a variant of the Zr2Fe12P7type.

Hypertrophic scars and keloids respond to dermal disruption with excessive collagen deposition and increased transforming growth factor (TFG)-β expression. Connective tissue growth factor (CTGF) is a downstream mediator of TGF-β activity that is associated with scar and fibrosis. The authors hypothesize that there is increased expression of CTGF by hypertrophic scar and keloid fibroblasts in response to TGF-β stimulation.

Discusses the video borescope usage in the nondestructive testing field. Features of video borescope; Evolution of video borescope digital features and system case technologies; Color of charge coupled device (CCD).

Human processed lipoaspirate PLA cells are multipotent stem cells, capable of differentiating into multiple mesenchymal lineages bone, cartilage, fat, and muscle. To date, differentiation to nonmesodermal fates has not been reported. This study demonstrates that PLA cells can be induced to differentiate into early neural progenitors, which are of an ectodermal origin. Undifferentiated cultures of human PLA cells expressed markers characteristic of neural cells such as neuronspecific enolase NSE, vimentin, and neuronspecific nuclear protein NeuN. After 2 weeks of treatment of PLA cells with isobutylmethylxanthine, indomethacin, and insulin, about 20 to 25 percent of the cells differentiated into cells with typical neural morphologic characteristics, accompanied by increased expression of NSE, vimentin, and the nervegrowth factor receptor trkA. However, induced PLA cells did not express the mature neuronal marker, MAP, or the mature astrocyte marker, GFAP. It was also found that neurally induced PLA cells displayed a delayedrectifier type Kcurrent an early developmental ion channel concomitantly with morphologic changes and increased expression of neuralspecific markers. The authors concluded that human PLA cells might have the potential to differentiate in vitro into cells that represent early progenitors of neurons andor glia. Plast. Reconstr. Surg.111 1922, 2003.

Fibroblast growth factors FGFs are a family of 21 cytokines with a broad spectrum of activities, including regulation of cell proliferation, differentiation, and migration. The various FGFs bind to one or more of four different tyrosine kinase receptor types. FGFs 1, 2, 5, 7, and 10 are upregulated during adult cutaneous wound healing. However, the expression of FGFs during fetal skin development and scarless wound healing has not been characterized. It was hypothesized that differential expression of FGF isoforms and receptors occurs during fetal skin development and that this differential expression pattern may regulate the transition from scarless repair to healing with scar formation.

Plastic surgery residency program directors are frequently interested in predictors of future career direction in their applicants. Many programs strive to train leaders in academic plastic surgery. To determine what factors may predict the ultimate fate of graduating plastic surgery residents, the authors reviewed the application files of 33 former residents from a single, major plastic surgery training program. The data from 29 residents were available for analysis. Nearly half of the residents graduating from the plastic surgery training program went into private practice. Two factors, the number of years taken off for research before entering the plastic surgery residency and the presence of children, were found to be indicative of a candidate's future career path. Of particular note, there was no difference between academic graduates and nonacademic graduates with regard to their intentions in their letters of recommendation and personal statements. This information is useful to both academic program directors and resident applicants. Plast. Reconstr. Surg.111 981, 2003.

Mg15Ir5Si2(tetragonal, P42/n, a = 1371, 7(1) pm, c = 873, 0(2) pm, Z=4, 1497 Reflexe, 103 Parameter, R1 = 0, 048) wurde durch Reaktion der Elemente bei 900 °C in verschweißten Tantalampullen dargestellt. Die Verbindung hat von allen bisher bekannten ternären Siliciden den höchsten Gehalt an Erdalkalimetall. Sie ist das erste Beispiel für ein Silicid mit einer isolierten Übergangsmetall‐Silicium‐Baugruppe, die in eine Matrix aus Nicht‐Übergangsmetallatomen eingelagert ist. Die Struktur enthält planare Ir2SiIrSiIr2‐Baugruppen, in denen die Siliciumatome annähernd trigonal planar von Iridiumatomen umgeben sind (dIr‐Si= 235 und 236 pm).

Human liposuction aspirates contain pluripotent adiposederived mesodermal stem cells that have previously been shown to differentiate into various mesodermal cell types, including osteoblasts and chondrocytes. To develop an autologous research model of bone and cartilage tissue engineering, the authors sought to determine whether rat inguinal fat pads contain a similar population of osteochondrogenic precursor cells. It was hypothesized that the rat inguinal fat pad contains adiposederived multipotential cells that resemble human adiposederived mesodermal stem cells in their osteochondrogenic capacity. To test this, the authors assessed the ability of cells isolated from the rat inguinal fat pad to differentiate into osteoblasts and chondrocytes by a variety of lineagespecific histologic stains.

The use of undifferentiated cells for cellbased tissue engineering and regeneration strategies represents a promising approach for skeletal muscle repair. For such strategies to succeed, a readily available source of myogenic precursor cells must be identified. We have previously shown that cells isolated from raw human lipoaspirates, called processed lipoaspirate cells, display multilineage mesodermal potential in vitro. Because human liposuctioned fat is available in large quantities and can be harvested with low morbidity, it may be an ideal source of stem cells for tissueengineering applications. In this study, processed lipoaspirate cells were isolated from raw lipoaspirates harvested from eight patients who underwent cosmetic surgery. Processed lipoaspirate cells were placed in promyogenic conditions for up to 6 weeks, and the expression of the myogenic markers MyoD1 and myosin heavy chain was confirmed by using structure, histology, and reverse transcriptasepolymerase chain reaction. Histologic results were quantitated as an indicator or myogenic differentiation levels. We found that induced human processed lipoaspirate cells form multinucleated cells after 3 weeks of induction, indicative of the formation of myotubes. In addition, MyoD1 and skeletal muscle myosin heavy chain are expressed at distinct time points during differentiation with MyoD1 expression preceding expression of myosin. Finally, approximately 15 percent of human processed lipoaspirate cells can be induced toward myogenic differentiation 6 weeks after induction. In summary, our findings suggest that human processed lipoaspirate cells differentiate into myogenic cells. Furthermore, these cells may be a useful source for skeletal muscle engineering and repair.

Fetal wounds pass from scarless repair to healing with scar formation during gestation. This transition depends on both the size of the wound and the gestational age of the fetus. This study defines the transition period in the fetal rat model and provides new insight into scarless collagen wound architecture by using confocal microscopy. A total of 16 pregnant SpragueDawley rats were operated on. Open fullthickness wounds, 2 mm in diameter, were created on fetal rats at gestational ages 14.5 days E14 n10, 16.5 days E16 n42, and 18.5 days E18 n42 term 21.5 days. Wounds were harvested at 24 n18 per gestational age and 72 hours n24 per gestational age. Skin at identical gestational ages to wound harvest was used for controls. The wounds were fixed and stained with hematoxylin and eosin, antibody to type I collagen, and Sirius red for confocal microscopic evaluation. No E14 rat fetuses survived to wound harvest. Wounds created on E16 fetal rats healed completely and without scarring. E16 fetal rat hair follicle formation and collagen architecture was similar to that of normal, nonwounded skin. Wounds created on E18 fetal rats demonstrated slower healing only 50 percent were completely healed at 72 hours compared with 100 percent of the E16 fetal rat wounds at 72 hours. Furthermore, the E18 wounds healed with collagen scar formation and without hair follicle formation. Confocal microscopy demonstrated that the collagen fibers were thin and arranged in a wispy pattern in E16 fetal rat wounds and in nonwounded dermis. E18 fetal rat wounds had thickened collagen fibers with large interfiber distances. Twomillimeter excisional E16 fetal rat wounds heal without scar formation and with regeneration of normal dermal and epidermal appendage architecture. E18 fetal rat wounds heal in a pattern similar to that of adult cutaneous wounds, with scar formation and absence of epidermal appendages. Confocal microscopy more clearly defined the dermal architecture in normal skin, scarless wounds, and scars. These data further define the transition period in the fetal rat wound model, which promises to be an effective system for the study of in vivo scarless wound healing.

Pseudoexfoliation syndrome (PEX) has been suggested to represent a blood-aqueous barrier impairment leading to a higher protein content in aqueous humour of eyes with PEX. However, the nature of a prospective PEX protein has not yet been described. We set out to re-evaluate protein content and examine protein composition for prospective PEX protein candidates in aqueous humour of eyes with PEX syndrome. Aqueous humour of 52 patients with PEX and 38 without PEX signs was sampled during cataract or glaucoma surgery. Total aqueous protein concentration in the samples was analysed in 43 PEX specimens and 32 non-PEX specimens according to Bradford. Aqueous protein composition of all samples was determined by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS PAGE) and silver staining. Screening for amyloids was performed in nine PEX samples and six non-PEX samples by Congo Red staining and polarised light microscopy. Aqueous protein concentration was not significantly increased in PEX eyes in comparison with non-PEX eyes. Furthermore, we could not detect any characteristic difference in protein band sizes of the two groups after SDS PAGE. However, we were able to show the presence of amyloid exclusively in aqueous humour of PEX patients. Conclusion:our results do not confirm a generally higher protein concentration in pseudoexfoliation syndrome eyes. This does not necessarily contradict a blood-aqueous barrier impairment but illustrates the variance in protein concentration between and within the two groups. No characteristic protein band allocatable to pseudoexfoliation syndrome proteins could be detected in any of the samples. However, our findings support the theory that the pseudoexfoliation syndrome is associated with an amyloid of a serum protein.

AbstractThe KRAB domain of human Kox1, a member of the KRAB C2H2 zinc finger family, confers strong transcriptional repressor activities even to remote promoter positions. Here, HDAC inhibitors were used to demonstrate that histone deacetylation is not required for mediating transcriptional repression of KRAB zinc finger proteins. Two reporter systems with either stably integrated or transiently transfected templates, both under control of strong viral promoters, were analyzed. Under all circumstances, HDAC inhibition did not alter the repression potential of the KRAB domain. In case of the stably integrated luciferase reporter gene system, neither expression levels of the KRAB fusion protein nor complex formation with its putative corepressor TIF1β were significantly changed. Furthermore, the TIF1β/KRAB complex was devoid of mSin3A and HDAC1. In the transient transfection system, the transcriptional repression induced by TIF1β and HP1α was not diminished by HDAC inhibitors, whereas the repressory activity of TIF1α was significantly affected. Thus, KRAB, TIF1β and HP1α are likely to be functionally linked. In conclusion, HDAC activity is not essential for the strong transcriptional repressor activity mediated by the KRAB domain of Kox1 in particular and, presumably, by KRAB domains in general. This feature might be helpful in identifying and characterizing target genes under the control of KRAB zinc finger proteins.

Protein kinase CK2 is a pleiotropic Ser/Thr kinase occurring as α2β2, α′2β2, or αα′β2tetramers. A requirement in serum‐stimulated cell cycle entry in both the cytoplasm and the nucleus of human fibroblasts for phosphorylation(s) by CK2 has been concluded from stimulation inhibition by microinjected antibodies against the regulatory subunit (β). We have now examined this idea more directly by microinjection‐mediated perturbation of phosphorylation and non‐phosphorylation interactions of the catalytic subunits (α and α′), and by verifying the supposed matching of the cellular partition of CK2 subunits in the fibroblasts employed. While immunostaining and cell fractionation indicate that the partitions of subunits indeed match each other (with their predominant location in the nucleus in both quiescent and serum‐stimulated cells), microinjection of substrate or pseudosubstrate peptides competing for the CK2‐mediated phosphorylation in vitro resulted in significant inhibition of serum stimulation when placed into the nucleus but not when placed into the cytoplasm. Also inhibitory were nuclear but not cytoplasmic injections of antibodies against α and α′ that affect neither their kinase activity in vitro nor their complexing to β. The data indicate that the role played by CK2 in serum‐stimulated cell cycle entry is predominantly nuclear and more complex than previously assumed, involving not only phosphorylation but also experimentally separable non‐phosphorylation interactions by the catalytic subunits.

The transconjunctival approach to the orbit is underutilized because of concern regarding inadequate exposure and higher postoperative rates of lower eyelid shortening and ectropion. All patients who had a transconjunctival incision performed for orbital surgery over the last 6 years 1990 to 1996 were studied. Patients who had a transconjunctival blepharoplasty were excluded. A total of 35 patients, average age 32 years, had 45 transconjunctival incisions performed. Lateral canthotomy or cantholysis was not done. Operations fell into three categories fracture plating alone, 10 22 percent splitcalvarial bone graft placement with or without plating, 26 58 percent and orbital decompression, 9 20 percent.

Antisense oligonucleotides are powerful tools for the in vivo regulation of gene expression. We have characterized the intracellular distribution of fluorescently tagged phosphorothioate oligodeoxynucleotides (PS-ONs) at high resolution under conditions in which PS-ONs have the potential to display antisense activity. Under these conditions PS-ONs predominantly localized to the cell nucleus where they accumulated in 20–30 bright spherical foci designated phosphorothioate bodies (PS bodies), which were set against a diffuse nucleoplasmic population excluding nucleoli. PS bodies are nuclear structures that formed in cells after PS-ON delivery by transfection agents or microinjection but were observed irrespectively of antisense activity or sequence. Ultrastructurally, PS bodies corresponded to electron-dense structures of 150–300 nm diameter and resembled nuclear bodies that were found with lower frequency in cells lacking PS-ONs. The environment of a living cell was required for the de novo formation of PS bodies, which occurred within minutes after the introduction of PS-ONs. PS bodies were stable entities that underwent noticeable reorganization only during mitosis. Upon exit from mitosis, PS bodies were assembled de novo from diffuse PS-ON pools in the daughter nuclei. In situ fractionation demonstrated an association of PS-ONs with the nuclear matrix. Taken together, our data provide evidence for the formation of a nuclear body in cells after introduction of phosphorothioate oligodeoxynucleotides.

The lack of scarring and fibrosis in healing fetal skin wounds may relate to a prolonged presence of hyaluronan (HA). It has been suggested that fetal wounds may lack hyaluronidase, but the hyaluronidase levels in fetal wounds remain unknown. The size of HA influences its biological action, especially in relation to angiogenesis, which is also reduced in fetal wound healing. The present study determined the levels and size of HA, as well as hyaluronidase levels, in fetal and adult lamb wounds. Wire mesh cylinders, or polyvinyl acetate sponges, were placed subcutaneously in fetal lambs at 75, 100 or 120 days gestation. Wound fluid and wound tissue were harvested 3, 7 or 14 days later. Samples were digested with papain and both HA and hyaluronidase activity were determined in a competitive ELISA assay. Size distribution of HA was estimated using a Sephacryl S1000 column and fractions were collected for HA determination. Adult wound fluid HA remained low (4–5 μg/ml) over the 14 days. Fetal fluids were similar on day 3, but increased to 15–25 μg/ml by day 7. In 75/100-day wounds, HA remained elevated at 14 days, but in 120-day fluids decreased to levels similar to adult fluid. The HA in all fluids was polydisperse with a main peak at 200 kDa. Hyaluronidase levels were detected in all samples, reaching a peak 7 days post-wounding. In adult wound fluids hyaluronidase was much higher than the fetal wound fluids. These data suggest that lower hyaluronidase levels in fetal wounds may underlie the different pattern of HA deposition seen in fetal wounds.

A system which allows sensitive and fast automated analysis of weakly labeled fluorescent specimens is described. It is tested in the analysis of c-fos expression stimulated by fetal calf serum and calibrated by quantitation of defined solutions injected into cells with the automated microinjection system. Low light level imaging technology combined with quantitative image processing methods and computer control of the hardware allows fully automated analysis of fluorescent molecules in single living or fixed cells. Reliable methods for subtraction of fluorescent background and automated identification of objects of interest in double-stained cells are described. The accuracy of quantitation is considerably improved by normalizing the fluorescence intensities of respective fluorophores in the same object by the method of ratio imaging. The error rate in determining the relative protein content in single cells is less than 15%. The method is applied to microinjection studies with a monoclonal antibody against casein kinase II subunit β. Microinjection of this antibody into synchronized cells specifically inhibits c-fos expression stimulated by fetal calf serum. In combination with the computer-automated capillary microinjection system, the technique will become a useful tool in experiments requiring quantitative single cell analysis. Copyright 1993, 1999 Academic Press

Generalized fin-lines are analyzed which consist of slots located on more than one interface between dielectric layers inserted in parallel with the E-plane of a waveguide. The analysis is based on a simplified formulation technique based on the spectral domain immittance matrix followed by a Galerkin's method. It allows as many slots as one desires at each interface, and the locations and the widths of these slots are arbitrary. The numerical process to carry out the present analysis is rather efficient. Some numerical data, including propagation constants of the propagating modes as well as characteristic impedances are presented for several structures considered useful for millimeter-wave circuit applications.

AbstractMactanamide (1), a new diketopiperazine containing the uncommon amino acid D-2,6-dihydroxyphenylalanine, has been isolated from the mycelium of an undescribed marine fungus of the genus Aspergillus. the new fungal strain was obtained from the surface of the brown marine alga Sargassumsp. collected near Mactan Island in the Philippines. the structure and absolute stereochemistry of 1was determined by interpretation of spectral data, by comparison with model compounds, and by measurement of the CD spectra of its component amino acids.

Mactanamide (1), a new diketopiperazine containing the uncommon amino acid D-2,6-dihydroxyphenylalanine, has been isolated from the mycelium of an undescribed marine fungus of the genus Aspergillus. the new fungal strain was obtained from the surface of the brown marine alga Sargassum sp. collected near Mactan Island in the Philippines. the structure and absolute stereochemistry of 1 was determined by interpretation of spectral data, by comparison with model compounds, and by measurement of the CD spectra of its component amino acids.

Dried suspensions ofPenicillium roquefortiThom, Coliphage T-1,Bacillus subtilisand tobacco mosaic virus were exposed to space on board the Gemini-IX-A and XII earth satellites and the Agena-VIII space rocket. All micro-organisms tested survived the direct exposure during the Gemini-IX-A experiment. In the Gemini-XII experiment only the T-1 phage survived the direct exposure. The survival was influenced by the suspending medium and depended on the species of the microorganism. After four months of space flight on the Agena-VIII space rocket surviving fractions between 2×10-3and 1.0 were found in the unopened flight container. However, micro-organisms exposed on the cover of the container during this period were completely inactivated. Shielding against solar ultraviolet radiation during flight resulted in survival of micro-organisms exceeding to that of the transport controls, and the survival was considered complete.

Survival of coliphageT1, tobacco mosaic virus (TMV) and spores ofPenicillium roquefortiThom after direct exposure to space on board 6 balloons, 6 sounding rockets and 3 satellites is related to the numbers of solar UV photons incident during exposure. The survival followed exponential curves leading to complete inactivation. Solar ultraviolet radiation of wavelengths 2000 Å to 3000 Å appears to be the main cause of inactivation of broth suspended phage and, probably, TMV.T1phage andPenicilliumspores prepared in saline were also affected by radiation shorter than 2000 Å, while TMV seems to be resistant to radiation of these wavelengths. The biological effectiveness of the solar spectrum in the area between 3000 Å to 50 000 Å was not significant. Sterilization of interplanetary spacecraft appears necessary since micro-organisms can easily be shielded against lethal radiation.

The phases of the cell cycle of Potorous tridactylus (PtK2) cells were determined in vitro by analysis of labeled mitoses for 37-½ hours after a tritiated thymidine pulse. The mean duration of DNA synthesis (tS) was 8 h. The mean duration of the gap phase before appearance of labeled mitoses was 5 h. Whereas the duration of the cell cycle (tC) based on analysis of labeled mitoses was 30 h, the doubling time (tD) derived from cell counts in the same cultures was only about 23 h. The analysis of the indices of labeled nuclei and mitoses suggests a stimulation of cells at the time of pulse labeling, which was maximal after beginning of the gap phase before DNA-synthesis, and possibly caused the observed difference between tCand tD.