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Large molecules can be generically separated from small ones, though partially and temporarily, in a pressure-driven flow inside a capillary. This transient incomplete separation has been only applied to species with diffusion coefficients different by at least an order of magnitude. Here, we demonstrate, for the first time, the analytical utility of transient incomplete separation for species with close diffusion coefficients. First, we prove

Molecular stream separation (MSS) is a promising complement for continuous-flow synthesis. MSS is driven by forces exerted on molecules by a field applied at an angle to the stream-carrying flow. MSS has only been performed with a 90° field-to-flow angle because of a rectangular geometry of canonic MSS; the second-order rotational symmetry of a rectangle prevents any other angle. Here, we propose a noncanonic circular geometry for MSS, which better aligns with the polar nature of MSS and allows changing the field-to-flow. We conducted in silico and experimental studies of circular geometry for continuous-flow electrophoresis (CFE, an MSS method). We proved two advantages of circular CFE over its rectangular counterpart. First, circular CFE can support better flow and electric-field uniformity than rectangular CFE. Second, the nonorthogonal field-to-flow orientation, achievable in circular CFE, can result in a higher stream resolution than the orthogonal one. Considering that circular CFE devices are not more complex in fabrication than rectangular ones, we foresee that circular CFE will serve as a new standard and a testbed for the investigation and creation of new CFE modalities.

Accurate Constant via Transient Incomplete Separation (ACTIS) is a new method for finding the equilibrium dissociation constant Kd of a protein-small molecule complex based on transient incomplete separation of the complex from the unbound small molecule in a capillary. This separation is caused by differential transverse diffusion of the complex and the small molecule in a pressure-driven flow. The advection-diffusion processes underlying ACTIS can be described by a system of partial differential equations allowing for a virtual ACTIS instrument to be built and ACTIS to be studied in silico. The previous in silico studies show that large variations in the fluidic system geometry do not affect the accuracy of Kd determination, thus, proving that ACTIS is conceptually accurate. The conceptual accuracy does not preclude, however, instrumental inaccuracy caused by run-to-run signal drifts. Here we report on assembling a physical ACTIS instrument with a fluidic system that mimics the virtual one and proving the absence of signal drifts. Furthermore, we confirmed method ruggedness by assembling a second ACTIS instrument and comparing the results of experiments performed with both instruments in parallel. Despite some unintentional differences between the instruments (caused by tolerances in sizes, positions, etc.) and noticeable differences in their respective separagrams, we found that the Kd values determined for identical samples with these instruments were equal. Conclusively, the fluidic system presented here can serve as a template for reliable ACTIS instrumentation.

For the first time, the long-term measurements of meteorological and oceanographic characteristics were measured using the anchored autonomous complex of the WaveScan buoy on the Southwestern shelf of the Peter the Great Bay (Japan Sea) from 21 April to 23 December 2016. The buoy anchor was set at 50 m depth. During 10 minutes of each hour, meteorological characteristics of the near surface layer of the atmosphere, sea water temperature at the horizon of 1.5 m, and vertical profiles of the current velocity vector from the near-surface layer to the near-bottom one were measured with resolution 4 m. During 20 minutes of each hour, the characteristics of waves on the sea surface were measured. In General, most of the meteorological characteristics measured on the buoy are in good agreement with the data of the NCEP-DOE AMIP-II, ERA-Interim and ERA5 reanalysis. The significant wave height for the whole frequency belt, mean wave period and mean spectral direction from the WaveScan Buoy measurements have the best statistical relationship, confidence level is 99%, with the correspondent significant height of the waves, mean wave period and mean spectral wave direction from reanalysis ERA5. The features of variability of vertical profiles of the current velocity vector on the synoptic and seasonal time scales that depend on wind speed and vertical stratification of density are determined. In the warm season the a significant left turn of the vector of the measured current velocity with depth is observed in the seasonal pycnocline below the surface layer of friction. At wind speed, not exceeding 5 m/s, the angle of the current velocity vector left turn reaches 170° in the bottom layer, where countercurrent is formed. During the passage of the Lionrock tropical cyclone, when and the daily mean wind velocity increase to 9 m/s, the angle of left turn of the current velocity vector in the pycnocline decreases to 20°. In the cold season a classic right turn of the current velocity vector is observed in the upper boundary layer, and the vertically average velocity vector within the 50 m layer, as well as the total drift flow in the Ekman friction layer, deviate 90° to the right from the surface wind velocity vector. The left turn of the current velocity vector at the buoy installation point was not observed in the cold season.

Analysis of cascading dynamic structure is carried out using measurements at three bottom autonomous stations installed on the continental slope of the Peter the Great Bay in the Sea of Japan. It is shown that in the course of immersion of surface waters disturbances of current velocity are generated. Interaction of temperature and current velocity fluctuations leads to fluxes of heat and momentum whose time-space and spectral structures is defined by duration and value of the immersed waters density anomalies. In the time of considerable anomalies, the immersion process has turbulent-vortical character.

ACTIS is a new method for finding the equilibrium dissociation constant Kd of a protein–small molecule complex based on transient incomplete separation of the complex from the unbound small molecule in a capillary. This separation is caused by differential transverse diffusion of the complex and the small molecule in a pressure-driven flow. The advection-diffusion processes underlying ACTIS can be described by a system of partial differential equations allowing for a virtual ACTIS instrument to be built and ACTIS to be studied in silico. The previous in-silico studies show that large variations in the fluidic system geometry do not affect the accuracy of Kd determination, thus, proving that ACTIS is conceptually accurate. The conceptual accuracy does not preclude, however, instrumental inaccuracy caused by run-to-run signal drifts. Here we report on assembling a physical ACTIS instrument with a fluidic system that mimics the virtual one and proving the absence of signal drifts. Furthermore, we confirmed method ruggedness by assembling a second ACTIS instrument and comparing the results of experiments performed with both instruments in parallel. Despite some differences between the instruments and, accordingly, significant differences in their respective separagrams, we found that the Kd values determined for identical samples with these instruments were equal. Conclusively, the fluidic system presented here can serve as a template for reliable ACTIS instrumentation.

The authors develop a new construction, manufacturing technology and method of calculating three-layer wall panels with heat-insulating packages embedded in a contour frame of lightweight concrete for agricultural and industrial buildings. The proposed technical solution gives the wall panels a number of significant distinguishing properties: the outer and inner plates of the heat-insulating package of the panel along the contour are embedded in the monolithic reinforced concrete frame; a layer of insulation is embedded in the contour frame, which ensures a decrease in its thermal conductivity; embedding of plates and insulation in the contour frame minimizes the labor capacity of the elements connection; placing a layer of insulation between two pre-made plates.

In familial cutaneous malignant melanoma (CMM), disruption of the retinoblastoma (pRB) pathway frequently occurs through inactivating mutations in the p16 (p16INK4A/CDKN2A/MTS1) gene or activating mutations in the G1-specific cyclin dependent kinase 4 gene (CDK4). Uveal malignant melanoma (UMM) also occurs in a familial setting, or sometimes in association with familial or sporadic CMM. Molecular studies of sporadic UMM have revealed somatic deletions covering the INK4A-ARF locus (encoding P16INK4Aand P14ARF) in a large proportion of tumours. We hypothesized that germline mutations in the p16INK4A, p14ARFor CDK4 genes might contribute to some cases of familial UMM, or to some cases of UMM associated with another melanoma. Out of 155 patients treated at the Institut Curie for UMM between 1994 and 1997, and interviewed about their personal and familial history of melanoma, we identified seven patients with a relative affected with UMM (n = 6) or CMM (n = 1), and two patients who have had, in addition to UMM, a personal history of second melanoma, UMM (n = 1), or CMM (n = 1). We screened by polymerase chain reaction single-strand conformation polymorphism the entire coding sequence of the INK4A-ARF locus (exon 1α from p16INK4A, exon 1β from p14ARF, and exons 2 and 3, common to both genes), as well as the exons 2, 5 and 8 of the CDK4 gene, coding for the functional domains involved in p16 and/or cyclin D1 binding. A previously reported polymorphism in exon 3 of the INK4A-ARF locus was found in one patient affected with bilateral UMM, but no germline mutations were detected, either in the p16INK4A, p14ARFor CDK4 genes. Our data support the involvement of other genes in predisposition to uveal melanoma. © 2000 Cancer Research Campaign

Introduction Les localisations digestives de lymphoproliferations indolentes T (LPI-T) decrites en 2000 par Ranheim et al., sont caracterisees par une colite recurrente, un infiltrat lymphoide T fait de cellules de taille moyenne, un index de proliferation bas et une evolution indolente (a la difference des lymphomes). Nous rapportons un cas avec atteinte buccale et digestive mimant une maladie de Crohn, d’evolution favorable sous traitement par cyclophosphamide. Observations Un homme ne en 1978, sans antecedent de maladie cœliaque, presentait depuis 2007 des ulcerations du palais et des gencives, tres invalidantes, entrainant une perte de poids. En 2008 douleurs abdominales et diarrhee revelaient une pan-colite, dont l’histologie consideree comme une colite ulcereuse conduisait a un traitement par adalimumab et mezalazine sans efficacite. Une nouvelle biopsie buccale realisee en 2014 et la relecture des lames de biopsie digestive et buccales anterieures mettaient en evidence un infiltrat T lymphocytaire fait de cellules monotones, de taille moyenne, CD2+, CD3+, CD5+, CD8+, TiA1+, granzyme B faible, CD56−, hybridation in situ EBER− avec une perte antigenique du CD7, un index de proliferation inferieur a 20 %, un clone T faisant porter le diagnostic de LPI-T. Le bilan ne montrait aucune autre localisation. Le patient etait traite par cyclophosphamide (150 mg/jour) avec une efficacite remarquable : disparition de la symptomatologie buccale et digestive, reprise de poids. Discussion La plus grande serie de LPI-T digestive rapportee dans la litterature comporte 10 cas avec une predominance masculine, un âge moyen au diagnostic de 48 ans, une atteinte predominante du colon et de l’intestin grele, un infiltrat fait de lymphocytes de taille moyenne le plus souvent CD8+ avec un index de proliferation bas, un clone T, et une evolution indolente. L’association a des ulcerations buccales est frequente (3/10 cas) et peut preceder l’atteinte digestive (13 ans dans un cas). Dans cette serie, aucune autre localisation de lymphome n’a ete decrite et sur les 38 mois de suivi moyen, aucune transformation en lymphome de haut grade n’a ete observee. L’errance diagnostique par meconnaissance a conduit a des traitements inadaptes (5 patients consideres comme des lymphomes agressifs et traites par polychimiotherapies, 2 patients ont eu des resections chirurgicales digestives, dont un une colectomie, pour un diagnostic de colite ulcereuse). Conclusion Des ulcerations buccales peuvent reveler une lymphoproliferation T indolente et preceder l’atteinte digestive. Ce tableau ne doit pas etre confondu avec une maladie de Crohn ou un lymphome agressif. Dans notre cas, le cyclophosphamide (150 mg/J) a entraine une disparition des lesions et une amelioration spectaculaire de l’etat general du patient.

Derivatives of the cyclodextrins, αCD, βCD, and γCD, in which all primary hydroxyls are substituted by amine pendant groups, may be synthesized efficiently from the per-6-bromo-6-deoxy-CD derivatives by direct reaction with amines. These ACD derivatives, which bear six, seven, or eight amine pendent groups, represent interesting biomimetic receptors and catalysts. The synthetic strategy relies on quantitative transformation and efficient purification as is demonstrated by preparation of 11 homogeneous ACD derivatives. The limitations of the synthesis and potential adaptations are illustrated by the synthesis of several more ACD derivatives to >95% purity. A synthetic route to a CD persubstituted with primary amine functionalities at the primary face, per-6-(aminomethyl)-6-deoxy-CD, yields an alternative reagent to the simple per-6-amino-6-deoxy-CD, which is more suitable for further synthetic transformations. The synthetic strategy is further adapted to preparation of a prototypical (6 + 1)-ACD derivative...

A semiempirical MNDO calculation was carried out for the singlet potential energy surface for the dimerization of acetonitrile oxide leading to dimethylfuroxane. The dimerization of nitrile oxides proceeds by a two-step mechanism. The rate-limiting step is formation of a dinitrosoethylene intermediate. A semiempirical AMI calculation was carried out to study the effect of phosphoryl substituents on formation of the transition state of the rate-limiting step in the dimerization of dimethoxyphosphorylnitrile oxide. Independently of the initial orientation of the two phosphorylnitrile oxide molecules, a gauche-oriented dimerica species is formed in the first step in the dimerization. There is hardly any specific effect ofthe phosphoryl substituents in the first step of phosphorylnitrile oxide dimerization.

3-Phosphotylated isoxazoles and isoxazolines react with nucleophiles either with cleavage of the P-C bond and subsequent ring opening or with retention of the heterocyclic ring depending on the structure of the ring substituents, nature of the nucleophile, and reaction conditions. This permits the selective chemical modification of the starting compounds.

Ninety-three collections of leucocytes by cytapheresis followed by separation of monocytes by centrifugal elutriation were undertaken in twelve metastatic cancer patients (four melanomas, six colon carcinomas, one ovarian carcinoma, and one lung cancer). The leucaphereses were performed aiming to collect a product, ready for introduction into the elutriation chamber, i.e., with low contamination by erythrocytes and granulocytes. The median collection of leucocytes was 7.3 x 10(9). After elutriation, purified monocytes (mean: 0.91 x 10(9)) were cultured with 3-5% autologous serum for 7 days in the presence of 250 IU/ml of recombinant human gamma-interferon (Rh-IFN gamma) for the last 18 h of culture. The median number of activated macrophages (MAK) available for reinfusion was 2.4 x 10(8) for each culture. The phenotypes and the antitumoral potentiality of MAK cells were documented. Reinfusions performed i.v. or i.p. were well tolerated with no major side effects. No complete tumor response was obtained. One partial response and two stabilizations of the disease were observed in one melanoma and two colon carcinomas.

a clinical study of 14 patients presenting both malignant melanoma and HIV infection, and analysis of the literature to determine the frequency and specific features of this association.ten men and four women of median age 43 years were included. In 50% of cases, the primary melanoma consisted of spreading superficial melanoma with a mean Breslow thickness of 2.83 mm. In two cases, regional lymph node metastasis was discovered but with no primary melanoma being identified. HIV infection was already documented on diagnosis of melanoma in 11 cases, and it was discovered in three cases at the time of surgery for melanoma (treatment of the primary melanoma in two cases, and in one case, regional lymph node dissection two years after the initial diagnosis). Eight patients died within a mean period of 39 months, with melanoma being the cause of death in six cases. Following relapse of melanoma, the course of the disease was severe, with mean stage IV survival of 3.6 months. No response to chemotherapy was observed where such treatment was feasible.the presence of HIV appears to be an aggravating factor for the outcome of metastatic melanoma.our study suggests the importance of clinical examination of pigmented lesions in HIV patients in order to ensure early identification of melanoma.

Androgen-producing tumors of the ovary are rare in postmenopausal women and are revealed by severe virilization. Leydig hilus cell tumors are the most frequent postmenopausal virilizing tumors. In this report, an unusual and rare cause of alopecia due to Leydig cell hyperplasia within the wall of a simple cyst and in the ovarian hilus is described.An 80 year-old woman complained of a 10-year history of severe androgenic alopecia associated with very mild facial hirsutism, without others signs of virilization. Hormonal blood levels showed markedly elevated testosterone. Computed tomographic scan of the adrenals and the ovaries revealed an enormous left ovarian cystic mass. Bilateral hystero-ophorectomy was performed. Histological examination demonstrated bilateral Leydig cell hyperplasia within the wall of the cyst and in the right ovarian hilus. Two months postoperative hormonal evaluation demonstrated dramatically decreased plasma levels of testosterone.The clinical, X ray and histologic aspects of this case, although rare, show that the presence of virilization should lead to a search for an androgen-secreting ovarian or adrenal tumor.

Diagnosis of pigmented skin tumors is often difficult. The aim of the present study is to evaluate high frequency ultrasound and Doppler ultrasound in the localization and the characterization of these tumors.One hundred and seventy six pigmented skin tumors including melanocytic and non melanocytic, benign and malignant lesions have been examined before resection using high frequency ultrasound (20MHz probe) and Doppler ultrasound (13MHz probe-Doppler frequency: 7MHz). Imaging data have been correlated with histological data, available in all cases.Tumors seen at ultrasound were hypoechoic lesions, some of which, particularly malignant melanocytic tumors were vascularized. Precise ultrasonographic location of the tumor was correlated to its histological location: epidermis for purely epidermal proliferations (seborrheic keratoses), superficial dermis for dermal proliferations originating from epidermis (benign dermal nevi, melanomas during vertical growth phase, basal cell carcinomas), middle and deep dermis for dermal proliferations originating from dermis (fibrous histiocytomas, angiomas).Ultrasound may help in assessing positive and differential diagnosis of pigmented skin tumors as it precises, before histological examination, the exact location of the lesion within the different skin layers and its origin. As far as malignant melanocytic tumors are concerned, high frequency ultrasound and Doppler ultrasound appear promising in diagnosis algorithm and pronosic of these lesions.

The purpose of the present study is to asses the value of high frequency ultrasonography (20 MHz), a new noninvasive imaging technique, in cutaneous tumors. Cutaneous tumors are clinically varied and their diagnosis is still based on histopathological analysis. 140 cutaneous tumors have been examined with US and imaging findings have been compared to the results of clinical and histological examinations. This study shows that high frequency ultrasonography, even though it cannot replace pathological analysis, may help the dermatologist in the positive diagnosis of some cutaneous tumors and in accessing the location and the in depth-extension of the lesions in the different layers of the skin.

gp130 acts as a common transducing signal chain for all receptors belonging to the interleukin (IL)-6 receptor family. The IL-6-related cytokines [IL-6, IL-11, oncostatin M (OSM), leukemia inhibitory factor, ciliary neurotrophic factor, and cardiotrophin] often modulate tumor phenotype and control the proliferation of many tumor cell lines. We demonstrate that melanoma cell lines release, in vitro and in vivo (when transplanted in nude mice), soluble gp130 (sgp130), a potential antagonist of cytokines from the IL-6 family. Biochemical analysis revealed that sgp130 derived from melanoma patients' sera or from culture supernatants of melanoma cell lines is a Mr 104,000 protein that resolved after deglycosylation as a Mr 58,000 protein. PCR and Northern blot analysis only identified one gp130 membrane mRNA, suggesting that the soluble form of gp130 is generated by proteolytic cleavage. OSM reproducibly increases sgp130 released by melanoma cell lines, whereas leukemia inhibitory factor stimulates the production of sgp130 in only one of three cell lines tested. This tumor-derived sgp130 is functional because it binds in solution to the IL-6-soluble IL-6 receptor (gp80) complex. Recombinant sgp130 inhibits the growth inhibitory activity of the IL-6-soluble IL-6 receptor complex and OSM on some melanoma cell lines. Therefore, this soluble gp130 represents a natural antagonist of cytokines from the IL-6 family.

In order to test the hypothesis that a combination of protease inhibitors with nucleoside analogues-agents known to inhibit different steps of the human immunodeficiency virus (HIV) life cycle--is likely to prove more effective in reducing viral loads than either of those modalities alone, we performed a 60 week, open-label trial in 32 HIV-positive patients with depressed CD4 T lymphocyte cell counts but no active AIDS-defining illnesses. For the first 2 weeks, patients received 600 mg twice daily of liquid ritonavir, a protease inhibitor; then zidovudine 200 mg three times daily and zalcitabine 0.75 mg three times daily were added to the treatment regimen. Mononuclear blood cell fractions were analysed for infected cell levels, using a co-culture system. HIV-1 RNA in plasma was measured both by reverse transcriptase-polymerase chain reaction (RT-PCR) and reverse transcriptase quantitative PCR (QcRT-PCR); lymphocyte counts were determined by standard laboratory methods. In the 2 weeks of ritonavir therapy, both the mean count of infectious blood cells and plasma HIV RNA levels decreased dramatically. Mean CD4 cell counts increased from 173 cells/mm3 at baseline to 286 cells/mm3; CD8 cell counts rose from 951 cells/mm3 to 1,141 cells/mm3. With the introduction of the nucleoside analogues, infectious cell counts and plasma virus dropped another log unit to a nadir at 8 weeks, while CD4 T lymphocyte counts continued to rise slowly. By week 28, 12 patients had withdrawn due to adverse events, none of which were life-threatening. At week 36, infectious material could not be detected in the cells of 10 of the 17 remaining patients; by week 60, four of the seven patients with residual viraemia at week 24 had undergone viral relapse. After the introduction of a more palatable capsule formulation of ritonavir at week 52, infectious cells and plasma virus were undetectable in 50-60% of patients. The combination of protease inhibitors and nucleoside analogues significantly reduces HIV load, and in some patients may suppress viral activity for sustained periods.

We have shown that chloronitroacetyl chloride (I), which is readily obtained by the nitration of trichloroethylene using HNO3-H2SO 4 as a nitrating mixture [2], may be a source of chloronitrile oxide. Carrying out the reaction of (I) with a twofold excess of monosubstituted acetylenes (II) in methylene chloride in the presence of an equimo!ar amount of sodium bicarbonate gave the corresponding 5-substituted 3-chloroisoxazoles (III) in 30-45% yield.

In a series of 12 patients suffering from an HIV infection, including 9 with confirmed AIDS, who complained about pharyngeal pain in a fixed site, having a progressive intensity and not relieved by antalgics and the specific treatments that were prescribed empirically, and for whom etiological investigation yielded negative results, Thalidomide proved to be the only effective means of healing the exulcerated, nail-mark lesions or the ulcerated, budding, neoplastic-like lesions, and of completely suppressing pain.

2542 Background: Vaccination with dendritic cells (DC), designed to express tumor antigens, is considered to be a possible strategy to elicit anti-tumour immune responses. Effective vaccines that mediate clinical responses in cancer patients may require generation of broad specific cytotoxic T lymphocytes directed against multiple epitopes. Allogeneic lysates from tumor cell lines are an attractive source of antigens as they contain multiple known and unknown tumor associated antigens (TAA). Methods: This is a multicentre, randomised, blinded phase I/II study in patients (pts) with stage IV (AJCC) metastatic melanoma. Safety was the primary objective; immune response (IR) and clinical response, using RECIST criteria, were secondary objectives. Patients were randomised to receive either non matured or matured DC loaded with lysate from 3 melanoma cell lines. DC were obtained from monocytes cultured with IL-13 and GM-CSF and, as required, matured with a bacterial extract and IFN-γ. Six immunizations of ∼2.0...

DNA sequences closely related to herpesvirus-like sequences have been found in AIDS-associated Kaposi's sarcoma. Using PCR, we found herpesvirus-like DNA sequences in Kaposi's lesions and normal adjacent skin in five patients with Mediterranean Kaposi's sarcoma. We did not find these sequences in tissues from patients without Kaposi's sarcoma. Semi-quantitative PCR revealed many more herpesvirus-like sequences in Kaposi's lesions than in unaffected skin. Our results reinforce the hypothesis that an infectious agent closely related to gamma-herpesvirus is implicated in the pathogenesis of Mediterranean and AIDS-associated Kaposi's sarcoma.

Human herpesvirus 8 (HHV-8) is a new virus which has been reported in Kaposi's sarcoma and some lymphoproliferative disorders such as Castleman's disease and body-cavity-based lymphoma. Because HHV-8 shares homology with Epstein-Barr virus (EBV), we searched for the presence of HHV-8 DNA sequences in various cutaneous T- and B-cell lymphoma by the polymerase chain reaction (PCR). Forty-seven HIV-negative patients with cutaneous lymphoma or large plaque parapsoriasis were enrolled in the study. For the detection of HHV-8 DNA sequences we used PCR followed by a hybridization with a digoxigenin-labelled probe and nested-PCR. HHV-8 DNA sequences could only be detected in a patient with large plaque parapsoriasis. Our study does not suggest any direct implication of HHV-8 in the pathogenesis of most cutaneous lymphoma. Serological studies will be helpful to appreciate if there is an epidemiological link between HHV-8 and cutaneous lymphomas.

We describe five cases of haemolytic uraemic syndrome (HUS) associated with HIV infection. HUS revealed HIV infection in one case. Three patients had AIDS. One was stage IV C2. HUS must be known as a complication of HIV infection.

Some patients with von Willebrand's disease do not respond to stimuli such as venous occlusion and infusion of a vasopressin analogue DDAVP. In these patients, fibrinolytic activity is not enhanced and von Willebrand's factor is not released into the blood. Skin biopsies and cryostat sections were used to study the fibrinolytic activity of skin vessels and localization of tissue plasminogen activator (t-PA) in three patients with severe form of von Willebrand's disease. On fibrin films, no fibrinolysis developed around the skin vessels of the patients; however, using specific polyclonal and monoclonal antibodies to t-PA, and peroxidase coupled specific IgG, presence of t-PA antigen was demonstrated in endothelial cells (EC) of all of them. In plasma no t-PA activity was detected either before or after venous occlusion although t-PA inhibitor activity was in a normal range. Small amounts of t-PA antigen was measured in blood by ELISA. From these results, it is concluded that in patients with severe forms of von Willebrand's disease, t-PA present in EC is not functional and can not transform plasminogen into plasmin.

The reaction of 1-acetyl-2-phenyl-4-methylpyrazolidine with imminium salts leads to the formation of 3-methyl-1,2,3,4-tetrahydropyrimido[1,2-a]-10-methylenedimethylimininium chloride, the alkaline hydrolysis of which gives 3-methyl-1,2,3,4-tetrahydropyrimido[1,2-a]-10-aldehyde. Similar reactions with 1-propionyl and 1-phenylacetyl derivatives leads to nonformylated indolization products. The relative reactivities of the imminium were studied.

To determine DNA-protein cross-linkages induced by mustine in cultures of human skin fibroblasts a radiometric method and fractionation of the cell lysate on hydroxyapatite were used. The formation of DNA-protein cross-linkages and their elimination during long-term culture of the cells after treatment with the mutagen were demonstrated.

The possibility of accumulation of cross-linkages in the DNA-protein complex was studied during stationary phase aging of cells in culture. Chinese hamster cells were used in the experiments, along with human fibroblasts. /sup 3/H-thymidine, /sup 14/C-valine, and /sup 14/C-leucine were added to the medium. The quantity of protein firmly bound with DNA was judged from the value of the coefficient /sup 14/C//sup 3/H determined with allowance for penetration of counting from the /sup 14/C-channel into the /sup 3/H-channel. The authors maintain that the results presented in this paper provide further evidence of the value of stationary phase cell cultures for the study of the mechanisms of aging and also of some of the general principles underlying hereditary pathology.

The effect of solubilization of deoxyribonucleoproteins (DNP) in a medium of near-physiological ionic strength after treatment with the mutagen N-nitroso-N-methylurea (NMU) is highly dependent on the NMU concentration. To convert DNP into a soluble state, the critical number of groups in the DNA and protein must evidently be modified. On the basis of data obtained by the circular dichroism method and by viscosimetry it is concluded that after treatment with NMU the DNP complex becomes soluble in solvents with near-physiological ionic strength largely as a result of labilization and dissociation of the DNA-protein bonds.