Avapritinib-based SAR studies unveil a binding pocket in KIT and PDGFRA.

Avapritinib is the only potent and selective inhibitor approved for the treatment of D842V-mutant gastrointestinal stromal tumors (GIST), the most common primary mutation of the platelet-derived growth factor receptor α (PDGFRA). The approval was based on the NAVIGATOR trial, which revealed overall response rates of more than 90%. Despite this transformational activity, patients eventually progress, mostly due to acquired resistance mutations or following discontinuation due to neuro-cognitive side effects. These patients have no therapeutic alternative and face a dismal prognosis. Notable, little is known about this drug's binding mode and its medicinal chemistry development, which is instrumental for the development of the next generation of drugs. Against this background, we solve the crystal structures of avapritinib in complex with wild-type and mutant PDGFRA and stem cell factor receptor (KIT), which provide evidence and understanding of inhibitor binding and lead to the identification of a sub-pocket (Gα-pocket). We utilize this information to design, synthesize and characterize avapritinib derivatives for the determination of key pharmacophoric features to overcome drug resistance and limit potential blood-brain barrier penetration.

Performance of a polymer coated silicon microarray for simultaneous detection of food allergen-specific IgE and IgG4.

BACKGROUND: Microarray-based component-resolved diagnostics (CRD) has become an accepted tool to detect allergen-specific IgE sensitization towards hundreds of allergens in parallel from one drop of serum. Nevertheless, specificity and sensitivity as well as a simultaneous detection of allergen-specific IgG4 , as a potential parameter for tolerance development, remain to be optimized. OBJECTIVE: We applied the recently introduced silicon chip coated with a functional polymer named copoly(DMA-NAS-MAPS) to the simultaneous detection of food allergen-specific IgE and IgG4 , and compared it with ImmunoCAP and ImmunoCAP ISAC. Inter- and intraslide variation, linearity of signal and working range, sensitivity and application of internal calibrations for IgE and IgG4 were assessed. METHODS: Native and recombinant allergenic proteins from hen's egg and cow's milk were spotted on silicon chips coated with copoly(DMA-NAS-MAPS) along with known concentrations for human IgE and IgG4 . A serum pool and 105 patient samples were assessed quantitatively and semi-quantitatively with the ImmunoCAP and ImmunoCAP ISAC and correlated with IgE- and IgG4 -specific fluorescence on silicon microarrays. RESULTS: Allergen-specific IgE and IgG4 were detected in parallel using two fluorescent dyes with no crosstalk. Results from the ImmunoCAP correlated better with microarray fluorescence than with ImmunoCAP ISAC except for the allergen ovomucoid. The working range of the silicon microarray for total hen's egg-specific IgE was comparable to the range of 0.1 to >100 kUA /L of the ImmunoCAP system, whereas for total cow's milk, the silicon microarray was less sensitive. Detectable allergen-specific IgG4 could be determined only for low concentrations, but still correlated positively with ImmunoCAP results. CONCLUSIONS: We confirmed the ability of the polymer coated silicon microarray to be comparably sensitive to the ImmunoCAP ISAC for various food allergens. This suggests that the copoly(DMA-NAS-MAPS) microarray is a low-cost, self-producible alternative to the commercial ImmunoCAP ISAC in allergy research.

Genetic aspects of cell line development from a synthetic biology perspective.

Animal cells can be regarded as factories for the production of relevant proteins. The advances described in this chapter towards the development of cell lines with higher productivity capacities, certain metabolic and proliferation properties, reduced apoptosis and other features must be regarded in an integrative perspective. The systematic application of systems biology approaches in combination with a synthetic arsenal for targeted modification of endogenous networks are proposed to lead towards the achievement of a predictable and technologically advanced cell system with high biotechnological impact.

Absolute beta-catenin concentrations in Wnt pathway-stimulated and non-stimulated cells.

The intracellular level of the proto-oncoprotein beta-catenin is a parameter for the activity of the Wnt pathway, which has been linked to carcinogenesis. The paper introduces a novel sandwich-based ELISA for the determination of the beta-catenin concentration in lysates from cells or tissues. The advantages of the method were proven by determining beta-catenin levels in cell lines and in cells after activation of the Wnt pathway. Analysis revealed high beta-catenin concentrations in the cell lines HeLa, KB, HT1080, MCF-7, U-87 and U-373, which had not been described before. Beta-catenin concentrations were compared in HEK293 and C57MG cells after activation of the Wnt pathway. The beta-catenin concentrations increased by different factors depending on whether the Wnt pathway was activated by incubation with LiCl or with Wnt-3a-conditioned medium. This finding indicated that the beta-catenin level depends on the way and level of Wnt pathway activation. The quantitative analysis of beta-catenin in colorectal tumours revealed high beta-catenin levels in tumours with truncating mutations in the APC gene.

[Accuracy of the autorefractor power refractor in clinical work--a comparative study]. / Messgenauigkeit des Autorefraktometers Power-Refractor in der klinischen Praxis -- eine vergleichende Studie.

BACKGROUND: The aim of this prospective study was to investigate accuracy and efficiency of the autorefractometer PowerRefractor compared to established autorefractometers and retinoscopy as standard method. PATIENTS AND METHODS: 150 patients (300 eyes) were examined. Patients were initially (pupil not influenced) analyzed with the Power Refractor (PR (neutral)). After that a examination followed under standardized Cyclopentolat cycloplegia with the Power Refractor (PR (cyclo)), the hand-held Retinomax K-plus (Nikon), the table-top mounted RK-5 (Canon) and retinoscopy. RESULTS: Deviation from retinoscopy (percentage of values with more than 1 dpt difference/maximal deviation): Sphere: PR (neutral): 66.3 %/8.5 dpt; PR (cyclo): 48 %/5 dpt; RK-5: 26.7 %/4.75 dpt; Retinomax K-plus: 6.7 %/1.25 dpt; Cylinder: PR (neutral): 12 %/6.25 dpt; PR (cyclo): 21 %/4.75 dpt; RK-5: 1.3 %/2.75 dpt; Retinomax K-plus: 2.3 %/2.5 dpt; spherical equivalent: PR (neutral): 44.7 %/7.1 dpt; PR (cyclo): 35.3 %/5.4 dpt; RK-5: 9 %/4.5 dpt; Retinomax K-plus: 1.3 %/1.,4 dpt; error of entire refraction: PR (neutral): 62.7 %/7.8 dpt; PR (cyclo): 61.3 %/7.3 dpt; RK-5: 24 %/4,5 dpt; Retinomax K-plus: 14.7 %/3.6 dpt CONCLUSIONS: The best accuracy compared to retinoscopy had Retinomax K-plus followed by RK-5. PowerRefractor had the biggest deviations of the tested devices. PowerRefractor may be a usefull device for screening small children and handicapped people because of the one meter observing distance. For a precise refraction especially in children a retinoscopy under cycloplegic conditions is still necessary.

Bioassays and selected chemical analysis of biocide-free antifouling coatings.

Over the years several types of biocide-free antifouling paints have entered the market. The prohibition of biocidal antifouling paints in special areas of some European countries such as Sweden, Denmark and Germany has favoured the introduction of these paints to the market. Several types of biocide-free antifouling paints were subjected to bioassays and selected chemical analysis of leachate and incorporated substances. Both non-eroding coatings (silicones, fibre coats, epoxies, polyurethane, polyvinyl) and eroding coatings (SPCs, ablative) were tested to exclude the presence of active biocides and dangerous compounds. The paints were subjected to the luminescent bacteria test and the cypris larvae settlement assay, the latter delivering information on toxicity as well as on efficacy. The following chemical analyses of selected compounds of dry-film were performed: The results of the bioassays indicated that none of the coatings analysed contained leachable biocides. Nevertheless, some products contained or leached dangerous compounds. The analyses revealed leaching of nonylphenol (up to 74.7 ng/cm2/d after 48 h) and bisphenol A (up to 2.77 ng/cm2/d after 24 h) from epoxy resins used as substitutes for antifouling paints. The heavy metal, zinc, was measured in dry paint film in quantities up to 576,000 ppm in erodable coatings, not incorporated as a biocide but to control the rate of erosion. Values for TBT in silicone elutriates were mostly below the detection limit of 0.005 mg/kg. Values for DBT ranged between <0.005 and 6.28 mg/kg, deriving from catalysts used as curing agents. Some biocide-free paints contained leachable, toxic and dangerous compounds in the dry film, some of which may act as substitutes for biocides or are incorporated as plasticizers or catalysts. Implications to environmental requirements and legislation are discussed.

Phenomenology and outcome of subjects with early- and adult-onset psychotic mania.

OBJECTIVE: This study examined clinical differences between subjects with early-onset and adult-onset psychotic mania. METHOD: Subjects were from an epidemiologically derived, hospitalized sample who met criteria for definite bipolar disorder after 24 months of follow-up and whose index episode had been manic. Information collected regarding demographic characteristics, psychotic and depressive symptoms, childhood behavior problems and school functioning, substance/alcohol use disorders, and episode recurrence for two subgroups were compared: those whose illness first emerged before age 21 (early onset) (N=23) and those whose first episode occurred after age 30 (adult onset) (N=30). RESULTS: A larger proportion of the early-onset subjects were male, had childhood behavior disorders, had substance abuse comorbidity, exhibited paranoia, and experienced complete episode remission less frequently during 24-month follow-up than the adult-onset subjects. CONCLUSIONS: These data add to the body of evidence that has suggested that many subjects with early-onset psychotic mania have a more severe and developmentally complicated subtype of bipolar disorder.

Determinants of dialyzer reuseability.

The authors observed a wide variability in the number of uses per dialyzer in their hemodialysis patients. In 54 patients who reused their dialyzers, heparin dose (U/kg/min) related directly and white blood cell count related inversely to reuseability. Average reuse was 5.1 +/- 2.9 (SD) in diabetics (n = 13) and 7.8 +/- 3.8 (SD) in non-diabetics (n = 41; p = 0.007). Of the diabetics, 77% achieved six or less reuses, and 59% of non-diabetics achieved six or more reuses (chi-square = 4.96; p less than 0.05). In non-diabetics, heparin dose was the most significant determinant of reuse, and in diabetics the major determinant was white blood cell count. Hematocrit levels, platelet count, erythropoietin use, or type of membrane (polysulfone/cellulose acetate) did not correlate with reuseability. It was concluded that reuseability of dialyzers is less in diabetic patients, patients on lower heparin doses (U/kg/min), and patients with higher white blood cell counts. Determinants of dialyzer reuseability warrant further study.

[The progestasert system--a 24-month-long active spiral. Clinical experiences with an intrauterine progesterone-releasing system]. / Das Progestasert-System--eine 24-Monats-Spirale. Klinische Erfahrungen mit einem intrauterinen Progesteron-freisetzenden System.

The Biograviplan (IUD) is a therapeutic system which, from a reservoir of 52 mg progesterone, permits the programmed release of the hormone within the uterus. It provides effective contraception over a period of 24 months. Spotting, intensified and prolonged menstrual bleeding can, in common with other IUDs, occur initially. Later, hypomenorrhoea and, occasionally, also silent menstruation extending over up to 3 cycles, can be seen. Dysmenorrhoea and menstrual pain are markedly reduced as compared with other IUDs. No intrauterine pregnancy was observed, but several extrauterine pregnancies occurred. Patients should be carefully selected for such a device, which can be recommended in particular for women with primary dysmenorrhoea or hypermenorrhoea.

[Management of pregnant women with thalassemia minor]. / Management bei Schwangeren mit Thalassaemia minor.

Owing to the fact that Germany has become the domicile of a great number of aliens from Mediterranean countries, an increasing number of thalassaemia cases have been reported and given cause for a thorough investigation. Since thalassaemia minor is a hereditary disease, a special prophylaxis has to take place in case of pregnancy: 1. examination of child's generator, 2. genetic counselling, 3. embryoscopy and extraction of embryonic blood, 4. if thalassaemia is diagnosed, an abortion should be taken into consideration.

[A report about 311 ovarian carcinomas, treated at the "Universitäts-Frauenklinik des Klinikums Mannheim der Universität Heidelberg" from 1960--1978 (author's transl)]. / Das Ovarialkarzinom an der Universitäts-Frauenklinik Mannheim. Eine Ubersicht über 311 Fälle.

In the medical histories no hints could be found, suggesting a definite risk-group for ovarian carcinoma. Symptoms and complaints are often uncharacteristic and no methods for early recognition are known. The therapy of the ovarian cancer is an operation, combined with chemotherapy, or radiation considering to the stage of the tumour. Most important for the prognosis is the degree of expansion, besides age of the patients and the histological pattern of the tumour. The three-years-survival rate of our patients with stage T1 is 71,7%, and with stage T4 only 13,5%. Above all advance in early diagnosis it is necessary to achieve higher survival rates.

[The prognosis of malignant ovarian tumours in correlation to the histological findings (author's transl)]. / Die Prognose der malignen Ovarialtumoren in Abhängigkeit vom histopathologischen Befund.

The prognosis of malignant ovarian tumours depends upon the stage at the onset of treatment when the histology is similar and in tumours of the same stage upon the histologic type of tumour. Ovarian cancers were histologically divided into 7 groups. The diagnosis in our series allowed to consider the following groups: Germinal cell tumours (group 1), tumours arising from isolated epithelial rests (group 2), tumours of follicular cell origin (group 3), tumours of the covering epithelium (group 4), and genetically indeterminate tumours (group 6). The stage T1 at the onset of treatment was most frequently diagnosed in tumours arising from follicular cells (61%). Tumours arising from the covering epithelium or genetically indeterminate tumours showed stage T1 in 15 or 16%. Secondary tumour at the onset of therapy was found in increasing frequency from group 1 to group 6. The survival rate decreases from group 1 to group 6. The worst prognosis show patients with genetically indeterminate tumours, followed by patients with malignant tumours of the covering epithelium. The best survival chance have patients with tumours of the germinal epithelium. It is therefore important to plan the post-operative treatment not only according to the stage of tumour but also according to the microscopic findings.

[Adhesion prophylaxis in surgical gynecology]. / Adhäsionsprophylaxe in der operativen Gynäkologie.

In operations near the internal genitalia it is important to prevent adhesions if fertility is to be preserved. As animal experiments and clinical experience have shown a fibrinolysin can be use for prevention of adhesions. In operations on the tube for restoring fertility and during chromolaparoscopy one should use a fibrinolysin, or example Varidase, and a cortisone derivative.

[Microscopic, micro-chemical and fluorescent optical findings in endometrium with a copper-T intrauterine device in situ (author's transl)]. / Histologische, histochemische und fluoreszenzoptische Befunde am Endometrium nach Einlage eines Kupfer-T-Pessars.

The uteri of ten patients who wore a copper-T for several months up to two years were examined immediately following hysterectomy by conventional microscopic techniques, fluorescent optical techniques after Acridin-Orange-Fluorochromization and by histo-chemical techniques for copper. With none of these methods changes in the endometrium could be found which explained the effect of the copper-T to inhibit implantation. Since practically any morphological change from the normal was absent it is suggested that the inhibiting effect on pregnancy of the copper intra-uterine devices is due to biochemical changes of the intra-uterine environment.

PIP: Sections of the uteri of 10 patients who had used IUDs for several months to 2 years were stained and treated by microchemical techniques to determine DNS, PNS, and copper levels. Examinations by microscope and fluorescent techniques showed no morphological effects of the copper on the endometrium. No deviations from normal DNS or RNS levels could be determined; thus the effectiveness of IUDs cannot be attributed to inhibition of estrogen or gestagen receptors. No elevated copper levels of the endometrium could be determined. It is concluded that the effectiveness of IUDs is due to a biochemical inhibition of implantation within the uterus.

[The treatment of pre-malignant hyperplasias of the endometrium with progesterone-carrying intra-uterine devices (author's transl)]. / Zur klinischen Anwendbarkeit gestagenbeladener Intrauterinpessare bei der Behandlung präkanzeröser Hyperplasien.

The local administration of progestional agents for the treatment of adenomatous hyperplasia of the endometrium is insufficient for two reasons: 1. The locally applied progestational agent does not reach the total surface of the endometrium especially not in the uterine fundus or in the tubal cornua. This is frequently due to uterine myomata and to the T-shape of the intra-uterine device which does not contain progestational agents within the T. 2. The progestational agent released by the intra-uterine device does not reach the deeper layers of the endometrium. It is therefore possible that the deeper layers of the endometrium undergo malignant change. At present it is therefore necessary to administer progestational agents in the treatment of adenotamous hyperplasias of the endometrium systemically.

[The effects and side-effects of the intra-cervical application of prostaglandin F-2 alpha during early pregnancy (author's transl)]. / Auswirkungen und Nebenwirkungen von intrazervikal appliziertem Prostaglandin F-2-alpha in der Frühschwangerschaft.

In order to avoid the traumatic dilatation of the cervix for therapeutic abortions 200 micrograms of prostaglandin F-2 alpha were injected into the cervix. The effect on dilatation of the cervix was compared with the injection of normal saline in the control group. Without side effects the injection of prostaglandin F-2 alpha resulted in dilatation of the cervix and softening of the cervix. Normal saline had no effect. At present it is unknown whether the dilatory effect of prostaglandin F-2 alpha on the cervix is local or secondary to uterine contractions.

PIP: Prostaglandin F2-alpha (200 mcg) was injected into the cervix to facilitate dilatation for first-trimester abortion. Patients included both nulliparae and multiparae in the 6th-11th week of pregnancy. 4 patients received injections of physiological saline solution; 4 others received 100 mcg injections; and 15 received 200 mcg of prostaglandin. Tokodynagraphs showed uterine contractions significantly increased in these last patients 7 hours after the injection; no change was noted in control patients. Patients receiving the smaller prostaglandin dose showed no effect. Regular labor contractions were not observed in any patient; the irregular contractions were easily inhibited with beta-sympathomimetics (Partusisten). 24 hours after the injections, in patients who had received the 200 mcg dose the cervix had opened to a Hegar 7, and could be dilatated to a Hegar 12 without difficulty.