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Background: Investigate retinal fluid changes via a novel deep-learning algorithm in real-world patients receiving faricimab for the treatment of neovascular age-related macular degeneration (nAMD). Methods: Multicenter, retrospective chart review and optical coherence tomography (OCT) image upload from participating sites was conducted on patients treated with faricimab for nAMD from February 2022 to January 2024. The Notal OCT Analyzer (NOA) algorithm provided intraretinal, subretinal and total retinal fluid for each scan. Results were segregated based on treatment history and fluid compartments, allowing for multiple cross-sections of evaluation. Results: A total of 521 eyes were included at baseline. The previous treatments prior to faricimab were aflibercept, ranibizumab, bevacizumab, or treatment-naive for 52.3%, 21.0%, 13.3%, and 11.2% of the eyes, respectively. Of all 521 eyes, 49.9% demonstrated fluid reduction after one injection of faricimab. The mean fluid reduction after one injection was −60.7nL. The proportion of eyes that saw reduction in fluid compared to baseline after second, third, fourth and fifth faricimab injections were 54.4%, 51.9%, 51.4% and 52.2%, respectively. The mean (SD) retreatment interval after second, third, fourth and fifth faricimab injection were 53.4 (34.3), 56.6 (36.0), 57.1 (35.3) and 61.5 (40.2) days, respectively. Conclusion: Deep-learning algorithms provide a novel tool for evaluating precise quantification of retinal fluid after treatment of nAMD with faricimab. Faricimab demonstrates reduction of retinal fluid in multiple groups after just one injection and sustains this response after multiple treatments, along with providing increases in treatment intervals between subsequent injections.

Objectives: Adaptive behavior, the ability to function in one’s environment, is compromised in individuals with Down syndrome (DS). Problems with adaptive behaviors may impact the individual’s physical and mental health-related quality of life (HRQOL). This study aims to describe the adaptive behavior of adults with DS and examine the role adaptive behavior has on their HRQOL. Methods: Sixty dyads of adults with DS and their caregivers completed the Vineland Adaptive Behavior Scales, Second Edition (Vineland-II) and QualityMetric 12-Item Short Form Health Survey, version 2 (SF-12v2). Results: All mean adaptive behavior levels were in the low range except coping and receptive communication, which were within the moderately low range. A significant negative correlation existed between age of the adult with DS and overall adaptive functioning, daily living skills, and socialization. Pairwise comparison revealed significant differences across Vineland-II domains, with ratings of socialization higher than daily living skills and communication, and daily living skills higher than communication. Only mental HRQOL was positively correlated with daily living skills, socialization, and overall adaptive functioning. Two parallel regression models were created, resulting in written communication (negative) and interpersonal relationships (positive) being the only significant predictors of mental HRQOL. Conclusions: Caregiver-reported adaptive behavior of adults with DS was in the low to moderately low ranges. By exploring associations between adaptive behavior and HRQOL, adults with DS, their caregivers, their providers (including healthcare, education, and service providers), and researchers can develop and target interventions to improve the adaptive behavior and, thus, the HRQOL of adults with DS.

The impact of ocean ecosystems on marine boundary layer aerosols and clouds has been the subject of much research but remains uncertain. Five experiments were recently conducted in the western North Atlantic to assess if the seasonally recurring phytoplankton bloom affects aerosol properties. These experiments include the second Western Atlantic Climate Study and four North Atlantic Aerosols and Marine Ecosystem Study cruises. Measurements of unheated and heated number size distributions, cloud condensation nucleus (CCN) concentrations, and aerosol composition were used to identify primary and secondary aerosol components that could be related to the state of the bloom. Only periods of clean marine air, as defined by radon, particle number concentrations, aerosol light absorption coefficient, and back trajectories, were included in the analysis. Nonvolatile material was found to be prevalent in the Aitken mode size range after heating to 230°, likely due to downward mixing from the free troposphere. CCN concentrations at 0.1% supersaturation were best correlated (r2= 0.73) with accumulation mode nss SO4=. Sea spray aerosol was only correlated with CCN during November when bloom accumulation had not yet occurred and dimethylsulfide concentrations were at a minimum. The fraction of CCN attributable to sea spray aerosol was less than 20% during March, May/June, and September, indicating the limited contribution of sea spray aerosol to the CCN population of the western North Atlantic atmosphere. The strongest link between the plankton bloom and aerosol and cloud properties appears to be due to biogenic non‐sea salt SO4=. Nonvolatile material in the Aitken mode after heating to 230 °C occurs in all seasons likely due to downward mixing from aloftNonsea salt sulfate links the phytoplankton bloom to cloud condensation nuclei (CCN) during bloom climaxSea spray aerosol only makes up a significant fraction of CCN in November when phytoplankton bloom accumulation has not begun

•Program theory of change must account for the lived experiences of medically and socially complex patients in order to affect dysfunctional patterns of acute care utilization.•Mental and emotional health, access to self-management resources, and patient-provider communication are key issues of importance to super-utilizing patients.•Transformation of didactic, patient education sessions to interactive, self-management support group sessions achieved success in improving patient engagement.•Lack of collaboration and compliance-oriented healthcare culture are primary threats to successful implementation of innovative healthcare delivery programs.•Linkage and navigation roles of healthcare staff are important in improving patient access to existing community resources, but further health system investments are required to effectively integrate community-based and social services into care delivery.•Peer support interventions are underutilized but hold great promise for addressing behavioral health needs of medically and socially complex patients.

Ein definitiver kurativer Therapieansatz ist mit einer operativen Tumorentfernung bisher nur für das lokal begrenzte Nierenzellkarzinom (NZK) vorhanden. Die Prognose beim metastasierten Nierenzellkarzinom (mNZK) ist vergleichsweise reduziert. Mit Einführung der zielgerichteten Therapie 2006 konnte eine deutliche Verbesserung des Gesamtüberlebens (OS) und des progressionsfreien Überlebens (PFS) erreicht werden. 2016 wurde mit der Zulassung des PD-1-Inhibitors Nivolumab das Therapiespektrum der Zweitlinie beim mNZK um eine neue Wirkstoffklasse erweitert. Zur Systemtherapie besteht weiterhin ein hoher therapeutischer Bedarf für zusätzliche Behandlungsoptionen, die einen relevanten Überlebensvorteil sowie ein anhaltendes und verbessertes Therapieansprechen ermöglichen. Die Systemtherapie für das mNZK umfasst aktuell insgesamt 11 verschiedene zielgerichtete Wirkstoffe aus 4 unterschiedlichen Substanzklassen. A definitive curative treatment exists only for localized renal cell carcinoma (RCC) by surgical resection. The prognosis for metastatic renal cell carcinoma (mRCC) is still comparatively limited. With the introduction of the targeted therapy in 2006, a significant improvement in overall survival (OS) and progression-free survival (PFS) was achieved. In 2016 the approval of the PD-1 inhibitor nivolumab enhanced the therapy spectrum of second-line treatment with a new class of active substances. For first-line treatment there is still a high therapeutic need for additional treatment options to achieve a relevant survival benefit and a sustained and improved response to treatment. The systemic therapy of mRCC currently comprises a total of 11 different targeted drugs from 4 different substance classes.

This study addressed three basic questions involving the amount of measurement needed to obtain valid estimates of reading growth. Participants were 41 students with learning disabilities from Grades 3 to 8, monitored across 10 weeks using curriculum-based measurement (CBM) of words read correctly (WRC). We compared growth slopes based on measurements taken weekly, every 2 weeks, every 3 weeks, every 4 weeks, and every 9 weeks to an estimate of “true slope.” Results showed that frequency of progress monitoring could be significantly reduced without detracting from the validity of growth estimates. However, validity was negatively affected by minimizing the number of WRC scores collected at each measurement occasion and by employing only one rather than four scores to estimate baseline.

Background: Selective inhibitors of cyclooxygenase‐2 (COX‐2) called coxibs, are effective anti‐inflammatory and analgesic drugs. Recently, these drugs were associated with an increased risk for myocardial infarction and atherothrombotic events. The hypothesis of thromboxane‐prostacyclin imbalance has been preferred to explain these unwanted effects. Methods: We studied the effects of 14 days intake of rofecoxib (25 mg q.d.), celecoxib (200 mg b.i.d.), naproxen (500 mg b.i.d.) and placebo in a randomized, blinded, placebo‐controlled study in young healthy volunteers (median age 25–30 years, each group n =10). We assessed prostanoid metabolite excretion (PGE‐M, TXB2, 6‐keto‐PGF1α, 11‐dehydro‐TXB2, 2,3‐dinor‐TXB2, and dinor‐6‐keto‐PGF1α), the expression of platelet activation markers (CD62P, PAC‐1, fibrinogen), platelet‐leukocyte formation, the endogenous thrombin potential, platelet cAMP content and plasma thrombomodulin level. Results: Naproxen suppressed biosynthesis of PGE‐M, prostacyclin metabolites and thromboxane metabolites and thrombomodulin levels. In contrast, both coxibs had an inhibitory effect only on PGE‐M, 6‐keto‐PGF1α, and on dinor‐6‐keto‐PGF1α, whereas TXB2, 2,3‐dinor‐TXB2and 11‐dehydro‐TXB2excretion were unaffected. None of the coxibs exerted significant effects on the expression of platelet activation markers, cAMP generation, platelet‐leukocyte formation, or on thrombomodulin plasma levels. Interestingly, platelet TXB2release during aggregation was enhanced after coxib treatment following arachidonic acid or collagen stimulation. Conclusion: In young healthy volunteers coxibs inhibit systemic PGE2and PGI2synthesis. Platelet function and expression of platelet aggregation markers are not affected; however, coxibs can stimulate TXB2release from activated platelets. Combined decrease in vasodilatory PGE2and PGI2together with increased TXA2in proaggregatory conditions may contribute to coxib side effects.

Abstract: Thin p-doped InGaN layers on p-doped GaN were successfully used to demonstrate a new type of low-resistance ohmic contact. A significant reduction of specific contact resistance can be achieved by increasing the free-hole concentration and the probability for hole tunneling through the Schottky barrier as a consequence of polarization-induced band bending. As obtained from the transmission-line method, the specific contact resistances of Ni (10 nm)/Au (30 nm) contacts deposited on InGaN capping layers were 1.210−2 Ωcm2 and 610−2 Ωcm2 for capping layer thicknesses of 20 nm and 2 nm, respectively.

Objective: Parameters of a pharmacokinetic-pharmacodynamic (PK-PD) model of levodopa have been claimed to reflect the magnitude of the dopaminergic deficit in patients with Parkinson's disease. The aim of this study was to correlate such parameters with positron emission tomography (PET) with levodopa tagged with 6-fluorine 18, an established imaging method for striatal dopaminergic neurons.Methods: Twenty-three patients in different disease stages (Hoehm and Yahr stage 2.5-5 [Hoehn MM, Yahr MD. Parkinsonism: onset, progression and mortality. Neurology 1967;4:427-42]; median duration, 12 years) were studied. PK-PD modeling followed a single oral dose of levodopa/benserazide. The sum score of the Columbia Rating Scale (CURSΣ) was used for clinical assessments. A nonparametric effect compartment approach assuming a sigmoidal Emaxmodel was applied to the PK-PD analysis of plasma levodopa concentrations and corresponding CURSΣ. Thereafter 6-[18F]levodopa PET was performed, and the influx rate constants (kc) for the putamen and the caudatus region were correlated with the median effective concentration (EC50) and the equilibrium half-life (Teq) of the PK-PD model.Results: (1) A significant correlation was observed between PK-PD parameters or with kcputamen as the dependent variable and the duration of the disease as the independent variable, which explains 33% of the variability of the EC50, 42% of the variability of Teq, and 36% of the variability of kc. (2) Significant correlations were observed between kcand either EC50or Teq, yielding the closest correlation for the putamen region (r = −0.47, P < .05; and r = 0.55, P < .01; respectively).Conclusions: Our findings show that key parameters of a PK-PD model of levodopa were in fairly close agreement with imaging of dopaminergic neurons by 6-[18F]levodopa PET. However, although PK-PD modeling of levodopa has been proven as a useful investigation of approaches aimed to restore dopaminergic deficits or to monitor disease progression, this modeling cannot serve as a pathomorphologic surrogate for the loss of striatal dopaminergic neurons.

By means of a dynamic acquisition protocol and a Fourier transform algorithm the scintigraphic technique can be used to assess gastric antral motility. In previous approaches, however, the definition of a region of interest (ROI) that gives rise to an unequivocal result of the analysis of the corresponding time-activity curve might be crucial. To overcome this problem we evaluated a new technique based on a fixed box-ROI and a simple weighting principle. Eight healthy volunteers ingested a meal that contained a 99Tcm-stannous colloid labelled omelette. Gastric emptying was assessed from static images taken at 30-mm intervals for 2 h. Anterior dynamic frames of 1 s each, which were acquired for 5 mm after each static acquisition, were used to determine the frequency of antral contractions. In order to allow for precise outlining of the antrum, each set of dynamic images was at first reframed into a single 5-mm image. The same ROI (20 pixels vertical, 15 pixels horizontal) was then positioned over the distal portion of the antrum on each reframed image. On each 1-s frame the obtained counts in each of the 15 columns of 20 pixels were summed. Then the localization of the geometric centre of radioactivity in the 15 columns was determined. On the assumption that the horizontal oscillations of the geometric centre of radioactivity in the ROI reflected the contractile activity of the antrum, the frequency of these contractions was calculated by fast Fourier transform analysis of the obtained time versus geometric centre curve. In all subjects, our technique gave unequivocal frequencies of antral contractions at each time point. Statistical analysis did not reveal any intraindividual variation in this frequency during gastric emptying. We believe that the simplified scintigraphic method is an advantageous alternative for assessing the frequency of antral contractions.

Metastatic progression of most common epithelial tumors involves a heterogeneous, transient loss of expression of the homotypic cell adhesion protein, E-cadherin, rather than the uniform loss of a functional protein resulting from coding region mutation. Indeed, whereas E-cadherin loss may promote invasion, reexpression may facilitate cell survival within metastatic deposits. The mechanisms underlying such plasticity are unclear. We now show that the heterogeneous loss of E-cadherin expression in primary human breast cancers reflects a heterogeneous pattern of promoter region methylation, which begins early prior to invasion. In cultured human tumor cells, such heterogeneous methylation is dynamic, varying from allele to allele and shifting in relation to the tumor microenvironment. Following invasion in vitro, which favors diminished E-cadherin expression, the density of promoter methylation markedly increased. When these cells were cultured as spheroids, which requires homotypic cell adhesion, promoter methylation decreased dramatically, and E-cadherin was reexpressed. These data show that the methylation associated with E-cadherin loss in human breast cancer is heterogeneous and unstable and suggest that such epigenetic plasticity may contribute to the dynamic, phenotypic heterogeneity that drives metastatic progression.

The effects of the dihydropyridine-type calcium antagonist (nitrendipine) and agonist (Bay K 8644) in comparison to atropine have been studied after intravenous administration on spontaneous and pelvic-nerve-induced contraction of rat urinary bladder. Bay K 8644 increased the basal internal bladder pressure as well as the amplitude of the spontaneous bladder contractions in a dose-dependent manner. In addition, an increase in systemic arterial blood pressure was noted for a period of about 20 min. In the presence of atropine the effects of Bay K 8644 on the urinary bladder were almost completely antagonized. Both nitrendipine and atropine reduced in a dose-dependent manner the amplitude of spontaneous and nerve-induced bladder contraction. The spontaneous and nerve-induced bladder contractions were significantly reduced by atropine or nitrendipine. Only nitrendipine caused a reduction of the spontaneous bladder contraction frequency. The systemic blood pressure was decreased significantly by nitrendipine but not after atropine administration. We suggest that both calcium antagonist and agonist can change the tension of the urinary bladder in vivo. As a side-effect the systemic blood pressure is altered. Atropine can antagonize the effect of BayK 8644 on the urinary bladder and reduces spontaneous and nerve-induced bladder contractions more specifically than nitrendipine.

Little is known about the important cellular substrates for protein kinase C and their potential roles in mediating protein kinase C-dependent processes. We evaluated the protein kinase C phosphorylation sites in a major cellular substrate for the kinase, a protein of apparent Mr80,000 in bovine and 60,000 in chicken tissues; we have recently determined the primary sequences of these proteins and tentatively named them the myristoylated alanine-rich C kinase substrates. The proteins were purified to apparent homogeneity from bovine and chicken brains, phosphorylated with protein kinase C, digested with trypsin, and the phosphopeptides purified and sequenced. Four distinct phosphopeptides were identified from both the bovine and chicken proteins. Two of the phosphorylated serines were contained in the repeated motif FSFKK, one in the sequence LSGF, and one in the sequence SFK. All four sites were contained within a basic domain of 25 amino acids which was identical in the chicken and bovine proteins. All of the sites phosphorylated in the cell-free system appeared to be phosphorylated in intact cells; an additional site may have been present in the proteins from intact cells. The identity of the phosphorylation site domains from two proteins of overall 65% amino acid sequence identity suggests a potential role for this domain in the physiological function of the myristoylated alanine-rich C kinase substrate proteins.

We treated 417 patients with upper ureteral stones with extracorporeal shock wave lithotripsy. All patients with obstructing stones underwent retrograde manipulation, which was successful in 57 per cent. Management of obstructing stones in situ (215 patients) with and without decompression of the collecting system required additional treatments in 13 per cent and ancillary procedures in 25 per cent. Nonocclusive ureteral stones were not manipulated. Treatment of these stones in situ slightly increased the need for postoperative ancillary procedures, compared to successful repositioning into the kidney (5.9 versus 3 per cent). Secondary treatments, however, were necessary as often as with occlusive stones. The main reason for failure of extracorporeal shock wave lithotripsy was the lack of fluid around an impacted stone. An energy absorptive effect of muscle tissue for stones projecting on the psoas muscle could not be demonstrated. The best and most consistent results were obtained when the stone was manipulated successfully into the renal collecting system.

Tension cracks are a major disturbance resulting from subsidence and differential settlement above underground coal mines. Recent engineering studies of subsidence indicate that cracks may close where tensile stresses causing the cracks are reduced or relaxed. This stress reduction occurs as mining in the area is completed. Crack closure was confirmed by a study in the Wasatch Plateau coal field of central Utah. Cracks occurred in both exposed bedrock and regolith in an area with maximum subsidence of 3 m. Mean closure rate was 0.3 cm per week with individual crack closure rates between 0.2 cm and 1.0 cm per week. The mean crack closure magnitude was 80% with closure magnitudes varying between 31% and 100%. Actual magnitude values ranged from 0.6 cm to 6.5 cm with a mean value of 3.8 cm. Statistical analysis compared width change status among cracks over time. It was found that: 1) a 41% probability existed that a crack would exhibit decreasing width per weekly measurement, 2) closure state sequences seem random over time, and 3) real differences in closure state sequence existed among different cracks. Les fissures d'extension constituent un grand dérangement de la surface de la terre; elles sont le produit du tassement et de l'affaissement différentiel sur les houillères souterraines. Des études récentes de l'affaissement indiquent que de certaines fissures peuvent se fermer en places où se réduisent ou se relâchent les forces de tension qui ont fait commencer ces fissures. Cette diminution de forces se produit pendant que s'achève l'exploitation des mines dans la région. On a confirmé certainement la fermeture des fissures pendant une étude du bassin houiller du Wasatch Plateau de l'Utah central. Des fissures se trouvaient et dans les roches de fond visibles et dans le régolite dans une région dont l'affaissement maximal est 3 m. La vitesse moyenne de constriction était 0.3 cm par semaine et la vitesse de la fermeture d'une fissure particulière était 80 pourcent et variait entre 31% et 100%. Les quantités véritables étaient comprises entre 0,6 cm et 6,5 cm, le moyen étant 3,8 cm. On a comparé, par l'analyse de la statistique, l'état changeant de la largeur parmi des fissures pendant le temps. On a trouvé (1) une probabilité de 41% qu'une fissure individuelle montrera quelque diminution de largeur par semaine; (2) une succession au hazard des états de fermeture sur une intervalle de temps; (3) des différences réelles entre les successions de l'état de fermeture des fissures différentes.

We evaluated 1,003 patients treated with extracorporeal shock wave lithotripsy after a mean followup of 19.1 months (range 12 to 26 months). Followup excretory urograms were normal in 97 per cent of the patients. Two-thirds of the patients reported further discharge of residual fragments, mainly during the first 3 months. Rehospitalization was necessary in 57 patients. Over-all, the rate free of stones after followup was 72.2 per cent and it was not different for primary and recurrent stone patients. Rates free of stones were influenced mainly by the primary stone location and the number of stones in a renal unit. Patients with lower caliceal stones had a rate without calculi of only 57.8 per cent. Almost identical results were obtained for stones other than in the lower calix, when fragments were found in the lower calix at the time the patient was discharged from the hospital. The pre-treatment stone volume, as determined by measuring the stone area in square millimeters, did not influence the final rates free of stones for calculi up to 400 mm.2, that is 2.4cm. of a sphere. Only calculi larger than 400 mm.2showed an inverse relationship to the final rate free of stones. Multiple stones yielded a success rate of 64 per cent, with 90 per cent of the patients having regrowth of residual fragments. Serious complications during followup were not encountered.

We examine roles of signaling by secreted ligands of the TGF-beta family during Drosophila oogenesis. One family member, the DPP ligand encoded by the decapentaplegic (dpp) gene, is required for patterning of anterior eggshell structures. This requirement presumably reflects the expression pattern of dpp in an anterior subset of somatic follicle cells: the centripetally migrating and the nurse cell-associated follicle cells. Similar requirements are also revealed by mutations in the saxophone (sax)-encoded receptor, consistent with the idea that DPP signaling is, at least in part, mediated by the SAX receptor. A loss of germline sax function results in a block in oogenesis associated with egg chamber degeneration and a failure of the transfer of nurse cell contents to the oocyte, indicating that TGF-beta signaling is required for these events. Some phenotypes of sax mutations during oogenesis suggest that SAX responds to at least one other TGF-beta ligand as well in the posterior follicle cells.

The myristoylated alanine-rich protein kinase C substrate (MARCKS) is a high affinity cellular substrate for protein kinase C. The MARCKS gene is under multiple modes of transcriptional control, including cytokine- and transformation-dependent, cell-specific, and developmental regulation. This study evaluated the transcriptional control of MARCKS gene expression during early development of Xenopus laevis. Xenopus MARCKS was highly conserved with its mammalian and avian homologues; its mRNA and protein were abundant in the maternal pool and increased after the mid-blastula transition (MBT). The Xenopus MARCKS gene was similar to those of other species, except that a second intron interrupted the 5'- untranslated region. By transiently transfecting XTC-2 cells and microinjecting Xenopus embryos with reporter gene constructs containing serial deletions of 5'-flanking MARCKS sequences, we identified a 124-base pair minimal promoter that was critical for promoter activity. Developmental gel shift assays revealed that a CBF/NF-Y/CP-1-like factor and an Sp1-like factor bound to this region in a manner correlating with the onset of Xenopus MARCKS transcription at MBT. Mutations in the promoter that abolished binding of these two factors also completely inhibited transcriptional activation of the MARCKS gene at MBT. The binding sites for these two factors are highly conserved in the human and mouse MARCKS promoters, suggesting that these elements might also regulate MARCKS transcription in other species. These studies not only increase our knowledge of the transcriptional regulation of the MARCKS genes but also have implications for the mechanisms responsible for zygotic activation of the Xenopus genome at MBT.

Abstract. The antiandrogen cyproterone acetate (CA), as well as oestrogens have been reported to influence pituitary-adrenal function in prostate cancer patients, but the clinical relevance of these findings is unknown. We therefore investigated serum cortisol (F), dehydroepiandrosterone sulphate (DS), testosterone (T) and prolactin (Prl) levels in patients treated with CA or oestradiol undecylate for at least 6 months. Hypothalamic-pituitary-adrenal function was further assessed by analysis of diurnal hormone variation and by ACTH stimulation and dexamethasone suppression tests. To differentiate between direct CA or oestrogen effects and secondary effects resulting from therapy-induced hypogonadism, we performed similar tests in untreated normogonadal and hypogonadal patients. CA treatment effected a significant decrease in serum F (−40%), DS (−73%) and T (−58%) levels and an increase in serum Prl (+118%). Oestrogen treatment resulted in markedly lowered T levels (−89%), slightly elevated serum F (+24%) and significantly increased serum Prl (+192%). Corresponding changes of F, DS and Prl could not be found in the untreated hypogonadal controls, thus indicating a direct drug-related effect. Neither diurnal rhythmicity of serum F nor adrenal response to ACTH stimulation or sensitivity to dexamethasone suppression significantly changed under CA or oestrogen treatment. We conclude that, although serum F levels may decrease under CA or increase slightly under oestrogen therapy for prostate carcinoma, these findings do not justify specific treatment, since neither clinical side effects nor an impairment of hypothalamic-pituitary-adrenal feedback occurs.

Promoter region CpG island methylation is associated with tumor suppressor gene silencing in neoplasia. GenBank sequence analyses revealed that a number of CpG islands are juxtaposed to multiple Alu repeats, which have been proposed as "de novo methylation centers." These islands also contain multiple Sp1 elements located upstream and downstream of transcription start, which have been shown to protect CpG islands from methylation. We mapped the methylation patterns of the E-cadherin (E-cad) and von Hippel-Lindau (VHL) tumor suppressor gene CpG island regions in normal and neoplastic cells. Although unmethylated in normal tissue, these islands were embedded between densely methylated flanking regions containing multiple Alu repeats. These methylated flanks were segregated from the unmethylated, island CpG sites by Sp1-rich boundary regions. Finally, in human fibroblasts overexpressing DNA methyltransferase, de novo methylation of the E-cad CpG island initially involved sequences at both ends of the island and the adjacent, flanking regions and progressed with time to encompass the entire CpG island region. Together, these data suggest that boundaries exist at both ends of a CpG island to maintain the unmethylated state in normal tissue and that these boundaries may be progressively overridden, eliciting the de novo methylation associated with tumor suppressor gene silencing in neoplasia.

Three technical modifications have been introduced into the Dornier HM-3 lithotriptor at the Marienhospital Herne: a low-pressure generator, which reduces the energy at the second focus by 30%, a modified semi-ellipsoid with an increased aperture and altered geometric dimensions, and the twin-pulse technique, releasing two impulses synchronized with a single R-wave. The treatment standard achieved with the conventional HM-3 device could be maintained, with a radiological success rate for stone disintegration of 95%. Procedural time could be reduced. The mean number of impulses and rate of second treatments increased, mainly due to the increasing proportion of ureteral and branched calculi. Cutaneous petechiae were rarely observed, suggesting less tissue damage with the lower pressures used. The profound reduction in pain perception with these technical modifications led to a new anesthesia technique. A combination of an oral anxiolytic and oral analgesic was effective in 94%. The results suggest that patient comfort and safety during ESWL treatment have been considerably improved.

The enantiomeric distributions of linalool and linalyl acetate in various natural products are measured by enantioselective gas chromatography on alkyl‐substituted cyclodextrins. Different plant cultivars were investigated by four extraction processes: steam distillation, solvent extraction, supercritical fluid extraction, and headspace analysis. Careful attention must be paid to linalool which undergoes partial racemization under certain processing conditions. In most cases, enantiomeric distribution is a powerful tool for authenticity testing.

The proteins necessary for signal transduction in cells responding to ligands of the TGF-beta family are largely unknown. We have previously identified Mad (Mothers against dpp), a gene that interacts with the TGF-beta family member encoded by decapentaplegic (dpp) in Drosophila. Assay of Mad's role in the DPP-dependent events of embryonic midgut development demonstrates that Mad is required for any response of the visceral mesoderm or endoderm to DPP signals from the visceral mesoderm. Replacement of the normal DPP promoter with a heterologous (hsp70) promoter fails to restore DPP-dependent responses in Mad mutant midguts. Experiments utilizing Mad transgenes regulated by tissue-specific promoters show that MAD is required specifically in cells responding to DPP. Immunohistochemical studies localize MAD to the cytoplasm in all tissues examined. Experiments in Xenopus embryos demonstrate that Drosophila MAD can function in the signaling pathway of BMP-4, a vertebrate homolog of dpp. Based on these results, we propose that Mad is a highly conserved and essential element of the DPP signal transduction pathway.

The Spemann organizer induces neural tissue, dorsalizes mesoderm and generates a second dorsal axis. We report the isolation and characterization of Smad10, which has all three of these Spemann activities. Smad10 is expressed at the appropriate time to transduce Spemann signals endogenously. Like the organizer, Smad10 generates anterior and posterior neural tissues. Smad10 appears to function downstream of the Spemann organizer, consistent with a role in mediating organizer-derived signals. Interestingly, Smad10, unlike previously characterized mediators of Spemann activity, does not appear to block BMP signals. This finding, coupled with the functional activity and expression profile, suggests that Smad10 mediates Spemann action in a novel manner.

Summary The influence of PGI2 and iloprost on the contents of high energy phosphates, cAMP, glycolytic metabolites and on cardiac performance was investigated in open-chest dogs subjected to 3 h of ischaemia, induced by occlusion of the left anterior descending coronary artery. The administration of drugs for 2h, starting 1h after the onset of ischaemia, resulted in a normalization and an appreciable reduction in the loss of high energy phosphates in the non — ischaemic and ischaemic left ventricular muscle, respectively, leading to an improvement in the phosphate potential of the cardiac tissue. A reduction in the ischemia-induced rise in cAMP levels was observed, indicating a lower catecholamine overflow in the presence of these drugs. PGI2 and iloprost in the dose employed significantly decreased cardiac preload, which was found to be elevated in the untreated animals after coronary artery ligation. The observed effect of PGI2 and its mimetic on the haemodynamics, as well as on myocardial cAMP and high energy phosphate levels of the heart, may be considered helpful in the phase of temporary imbalance between energy demand and supply during acute ischaemia.

In order to study cell tropism and attenuation of hepatitis A virus (HAV), the genome of HAV wild-type GBM and two cell culture-adapted variants, GBM/FRhK and GBM/HFS, were cloned and sequenced after amplification by reverse transcriptase-PCR. During virus cultivation, the HAV variant GBM/FRhK had a strict host range for FRhK-4 cells, in contrast to GBM/HFS, which can be grown in HFS and FRhK-4 cells. The HAV variant GBM/HFS was shown to be attenuated when inoculated into chimpanzees (B. Flehmig, R. F. Mauler, G. Noll, E. Weinmann, and J. P. Gregerson, p. 87-90, in A. Zuckerman, ed., Viral Hepatitis and Liver Disease, 1988). On the basis of this biological background, the comparison of the nucleotide sequences of these three HAV GBM variants should elucidate differences which may be of importance for cell tropism and attenuation. The comparison of the genome between the GBM wild type and HAV wild types HM175 (J. I. Cohen, J. R. Ticehurst, R. H. Purcell, A. Buckler-White, and B. M. Baroudy, J. Virol. 61:50-59, 1987) and HAV-LA (R. Najarian, O. Caput, W. Gee, S. J. Potter, A. Renard, J. Merryweather, G. Van Nest, and D. Dina, Proc. Natl. Acad. Sci. USA 82:2627-2631, 1985) showed a 92 to 96.3% identity, whereas the identity was 99.3 to 99.6% between the GBM variants. Nucleotide differences between the wild-type and the cell culture-adapted variants, which were identical in both cell culture-adapted GBM variants, were localized in the 5' noncoding region; in 2B, 3B, and 3D; and in the 3' noncoding region. Our result concerning the 2B/2C region confirms a mutation at position 3889 (C-->T, alanine to valine), which had been shown to be of importance for cell culture adaptation (S. U. Emerson, C. McRill, B. Rosenblum, S. M. Feinstone, and R. H. Purcell, J. Virol. 65:4882-4886, 1991; S. U. Emerson, Y. K. Huang, C. McRill, M. Lewis, and R. H. Purcell, J. Virol. 66:650-654, 1992), whereas other mutations differ from published HAV sequence data and may be cell specific. Further comparison of the two cell culture-adapted GBM variants showed cell-specific mutations resulting in deletions of six amino acids in the VP1 region and three amino acids in the 3A region of the GBM variant GBM/FRhK.

We isolated and sequenced a cDNA clone encoding the bovine "80- to 87-kDa" protein, a major cellular substrate for protein kinase C. An open reading frame of 1005 base pairs predicted a protein of 335 amino acids (Mr, 31,949). Despite this predicted size, the protein migrated on SDS/polyacrylamide gels with an apparent molecular weight of 80-87,000 after expression of the cDNA in cells lacking the protein. It was highly enriched in alanine (28.4 mol %), contained an amino-terminal myristoylation consensus sequence, and included a 25-residue basic domain containing the known protein kinase C phosphorylation sites. Two mRNA species (2.6 and 4.4 kilobases) were most highly expressed in brain, spinal cord, spleen, and lung, in parallel with the distribution of immuno-reactive protein. Genomic blot analysis indicated the likelihood of a single gene coding for this mRNA. We propose the name myristoylated alanine-rich C kinase substrate (MARCKS) for this protein.

Abstract: Concerning Medieval French, text-processing has to handle two major problems: first the under-developed state of linguistic knowledge of scholars; secondly the very large variations in forms and spellings occurring throughout the manuscripts. In order to extract and provide lexicographical and grammatical information from many yet undescribed texts, our research group has decided to devise an expert system which would help identify and parse the text-words and would be linked to an open relational database. The design of our expert system is outlined here: we especially outline the initial knowledge base and the nature of the different sets of rules we will make use of. Some examples describe how the system will operate and the types of advantages that are expected from its use. Finally the paper lists the studies and published work which in the last years have provided the groundwork for this project.

Models of transperitoneal urea transport are generally based on the one-compartment assumption, i.e. that the plasma water urea concentration in the peritoneal capillary bed is equal to the plasma water urea concentration in the peripheral veins. The aim of this study was to investigate the mechanism(s) of transperitoneal urea transport and to test the one-compartment assumption for urea. A total of 12 nested models were formulated and validated on the basis of experimental results obtained from 23 non-diabetic patients undergoing peritoneal dialysis. The validation procedure demonstrated that transperitoneal transport of urea probably involves diffusion, non-lymphatic convection and lymphatic convection. It was furthermore demonstrated that the inclusion of lymphatic convection changes the mass transfer area coefficient considerably. Finally, no deviation from the one-compartment assumption was demonstrated by our results.

We have evaluated the possibility that a major, abundant cellular substrate for protein kinase C might be a calmodulin-binding protein. We have recently labeled this protein, which migrates on sodium dodecyl sulfategel electrophoresis with an apparent Mrof 60,000 from chicken and 80,000-87,000 from bovine cells and tissues, the myristoylated alanine-rich C kinase substrate (MARCKS). The MARCKS proteins from both species could be cross-linked to 125I-calmodulin in a Ca2+-dependent manner. Phosphorylation of either protein by protein kinase C prevented 125I-calmodulin binding and cross-linking, suggesting that the calmodulin-binding domain might be located at or near the sites of protein kinase C phosphorylation. Both bovine and chicken MARCKS proteins contain an identical 25-amino acid domain that contains all 4 of the serine residues phosphorylated by protein kinase C in vitro.In addition, this domain is similar in sequence and structure to previously described calmodulin-binding domains. A synthetic peptide corresponding to this domain inhibited calmodulin binding to the MARCKS protein and also could be cross-linked to 125I-calmodulin in a calcium-dependent manner. In addition, protein kinase C-dependent phosphorylation of the synthetic peptide inhibited its binding and cross-linking to 125I-caImodulin. The peptide bound to fluorescently labeled 5-dimethylaminonaphthalene-1-sulfonyl-cal-modulin with a dissociation constant of 2.8 nM, and inhibited the calmodulin-dependent activation of cyclic nucleotide phosphodiesterase with an IC50of 4.8 nM. Thus, the peptide mimics the calmodulin-binding properties of the MARCKS protein and probably represents its calmodulin-binding domain. Phosphorylation of these abundant, high affinity calmodulin-binding proteins by protein kinase C in intact cells could cause displacement of bound calmodulin, perhaps leading to activation of Ca2+-calmodulin-dependent processes.

Regulation of phosphate uptake was studied in HeLa cell lines after transfection with DNA encoding the human 5-HT1Areceptor. Phosphate uptake was saturable and >90% sodium-dependent, with Vmax≈ 4.3 nmol/mg protein/3 min and Km≈0.4 mMphosphate. Activators of protein kinase A decreased Vmaxby ≈ 30-35% without changing Km. Treatment with 5-HT or the 5-HT1A-specific agonist 8-OH-2-(di-n-propylamino) 1,2,3,4-tetrahydronaphthalene increased Vmax≈ 40% without affecting Km. This effect was blocked by pretreatment with the 5-HT1antagonists, methiothepine and spiperone, or pertussis toxin. Surprisingly, the stimulation was not secondary to an inhibition of adenylyl cyclase because 5-HT stimulated phosphate uptake ≈ 20% in the presence of 1 mM8-Br-cAMP. Rather, the primary pathway linked to the stimulation of phosphate uptake involved activation of protein kinase C because (i) 5-HT measurably activated protein kinase C in these cells, (ii) activators of protein kinase C (phorbol esters and diacylglycerol analogues) stimulated phosphate uptake in these cells (iii) the half-maximal doses for 5-HT-induced phosphatidylinositol hydrolysis and stimulation of phosphate uptake were virtually equivalent, and both effects were equally sensitive to pertussis toxin, and (iv) the stimulation was markedly attenuated in cells made deficient in protein kinase C. These results demonstrate that the stimulation of phosphatidylinositol hydrolysis by the 5-HT1Areceptor can generate physiologically measurable effects on cellular transport and suggest that such accessory pathways may play a prominent role in signal transduction.

Radioisotope bone scanning is a sensitive indicator of metastatic disease. Localization of the hot spot for biopsy is difficult. Today, a hand-held gamma probe enables the surgeon intraoperatively to accomplish the localization procedure easily. As this technique is described in American literature exclusively, more recent European findings is presented here. During one month two patients were diagnosed with breast cancer. Bone scanning disclosed minor hot spots in the ribs. On the day of surgical biopsy the localization of the hot spots was guided by a gamma probe. For the biopsy a dermal punch biopsy needle was employed, and fine needle aspiration biopsies were also taken.The procedures were easy and completed within a few minutes. The post-operative courses were uneventful. In one patient, final histology showed malignancy in both biopsies. In the other patient, only the fine needle aspiration biopsy was malignant. The punch biopsy was abnormal, but did not contain malignant tissue.The gamma probe is an important tool in the performance of rib biopsies of nonpalpable lesions. The punch biopsy technique is simple and combined with fine needle aspiration biopsy the method has proven to be sensitive and accurate. The punch biopsy technique furthermore reduces the risk of postoperative pneumothorax.

A case of melanosis of the gall bladder is described. Ultrastructural examination confirms that the pigment is related to lipofuscin.