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The molecular nanoscale organization of the surfaceome is a fundamental regulator of cellular signaling in health and disease. Technologies for mapping the spatial relationships of cell surface receptors and their extracellular signaling synapses would unlock theranostic opportunities to target protein communities and the possibility to engineer extracellular signaling. Here, we develop an optoproteomic technology termed LUX-MS that enables the targeted elucidation of acute protein interactions on and in between living cells using light-controlled singlet oxygen generators (SOG). By using SOG-coupled antibodies, small molecule drugs, biologics and intact viral particles, we demonstrate the ability of LUX-MS to decode ligand receptor interactions across organisms and to discover surfaceome receptor nanoscale organization with direct implications for drug action. Furthermore, by coupling SOG to antigens we achieved light-controlled molecular mapping of intercellular signaling within functional immune synapses between antigen-presenting cells and CD8+ T cells providing insights into T cell activation with spatiotemporal specificity. LUX-MS based decoding of surfaceome signaling architectures thereby provides a molecular framework for the rational development of theranostic strategies., Nature Communications, 12 (1), ISSN:2041-1723

Background and importance We present the case of a young patient with T lymphoblastic lymphoma (T-LBL) who developed severe toxicity after receiving an anthracycline based protocol that is generally well tolerated by healthy patients. We assume that the substitution of daunorubicin by doxorubicin due to a nationwide shortage of daunorubicin played a crucial role in developing severe toxicity. Aim and objectives The 19-year-old man with no previous illnesses was diagnosed in December 2019 with T-LBL, a rare, aggressive neoplasm of precursor T cells that progresses rapidly and requires prompt diagnosis and medical intervention. T-LBL shows morphological and immunophenotypical similarities to acute lymphoblastic leukaemia (ALL). T-LBL treatment is the same as for ALL. Material and methods The induction protocol consisted of dexamethasone, vincristine, daunorubicin and pegasparaginase. Due to a nationwide shortage, daunorubicin (30 mg/m²) was substituted by doxorubicin (25 mg/m²). Results Chemotherapy was initially well tolerated, but beginning on day 15, the patient developed pronounced mucositis and increased skin toxicity (hand–foot syndrome, grade IV). Moreover, coagulation parameters deteriorated, and repeated transfusions with erythrocytes and platelet concentrates were needed. After administration of pegasparaginase on day 31, liver values increased, and finally, the patient had to be transferred to the intensive care unit due to fulminant pancreatitis. After 3 days, the patient could be transferred back to our ward. However, within 2 weeks, the patient developed sensory disturbances in all extremities, which was classified as chemotherapy associated polyneuropathy. In the further clinical course, the patient‘s general condition improved, and PET-CT showed complete metabolic remission. Due to the severe chemotherapy associated side effects, intensive consolidation treatment, according to the protocol, was cancelled. Instead, a consolidating therapy with nelarabine was carried out without complications. To date (October 2020), the patient is in a good clinical condition and has not developed disease recurrence. Probability assessment using the Naranjo algorithm resulted in ‘probable adverse drug reaction’ (score=6). Conclusion and relevance Our case report underlines the fact that shortages of essential anticancer drugs can have a particular impact on the efficacy and safety of established chemotherapy regimens, as these medicines often have few or no proven effective alternatives. References and/or acknowledgements Patel S, et al. A single-centre experience of the nationwide daunorubicin shortage: substitution with doxorubicin in adult acute lymphoblastic leukaemia. Leuk Lymphoma 2013;54:2231–5. Conflict of interest No conflict of interest

CD20 is a B cell-specific membrane protein and represents an attractive target for therapeutic antibodies. Despite widespread usage of anti-CD20 antibodies for B cell depletion therapies, the biological function of their target remains unclear. Here, we demonstrate that CD20 controls the nanoscale organization of receptors on the surface of resting B lymphocytes. CRISPR/Cas9-mediated ablation of CD20 in resting B cells resulted in relocalization and interaction of the IgM-class B cell antigen receptor with the coreceptor CD19. This receptor rearrangement led to a transient activation of B cells, accompanied by the internalization of many B cell surface marker proteins. Reexpression of CD20 restored the expression of the B cell surface proteins and the resting state of Ramos B cells. Similarly, treatment of Ramos or naive human B cells with the anti-CD20 antibody rituximab induced nanoscale receptor rearrangements and transient B cell activation in vitro and in vivo. A departure from the resting B cell state followed by the loss of B cell identity of CD20-deficient Ramos B cells was accompanied by a PAX5 to BLIMP-1 transcriptional switch, metabolic reprogramming toward oxidative phosphorylation, and a shift toward plasma cell development. Thus, anti-CD20 engagement or the loss of CD20 disrupts membrane organization, profoundly altering the fate of human B cells., Proceedings of the National Academy of Sciences of the United States of America, 118 (7), ISSN:0027-8424, ISSN:1091-6490

CD20 is a B cell specific membrane protein and a target of therapeutic antibodies such as rituximab (RTX). In spite of the prominent usage of anti-CD20 antibodies in the clinic little is known about the biological function of CD202. Here we show that CD20 controls the nanoscale organization of receptors on the surface of resting B lymphocytes. A CRISPR/Cas-based ablation of CD20 in Ramos B cells results in a relocalisation of the IgM B cell antigen receptor (IgM-BCR) and the co-receptor CD19. The resulting IgM-BCR/CD19 signaling synapse leads to transient B cell activation followed by plasma cell differentiation. Similarly to CD20-deficient Ramos cells, naive human B cells treated with rituximab in vitro or isolated from patients during rituximab administration display hallmarks of transient activation characterized by the formation of the IgM-BCR/CD19 signaling synapse, followed by CD19 and IgM-BCR downregulation. Moreover, increased expression of specific plasma cell genes can be observed after rituximab treatment in relapsed CLL patients. In summary we identify CD20 as a gatekeeper of the resting state on human B cells and demonstrate that a disruption of the nanoscale organization of the B cell surface via CD20 deletion or anti-CD20 treatment profoundly alters B cell fate.

Hydrodynamic forces on intertidal flats vary over a range of temporal and spatial scales. These spatiotemporal inhomogeneities have implications for intertidal flat morphodynamics and ecology. We determine whether storm events are capable of altering the long-term morphological evolution of intertidal flats, and unravel the contributions of tidal flow, wind-driven flow, waves, and water depth on inhomogeneities in bed level dynamics (bed level changes over ~days) across these areas. We complement decades of bed level measurements on eight intertidal flats in two estuaries in the Netherlands with an extensive 1-month field campaign on one of those flats. Across this intertidal flat, the hydrodynamics and morphodynamics of a storm event were captured, including the post-storm recovery. We show that individual events can persistently alter the morphological evolution of intertidal flats; magnitudes of some bed level changes are even comparable to years of continuous evolution. The morphological impacts of events are largely controlled by the relative timing of the forcing processes, and not solely by their magnitudes. Spatiotemporal variations in bed level dynamics of intertidal flats are driven by a combination of: (1) the inhomogeneous distributions of the hydrodynamic forcing processes (including the under-explored role of the wind); and (2) the linear proportionality between bed level dynamics and the local bed slope.

Reinforcement value enhancement by nicotine of non-nicotine rewards is believed to partially motivate smoking behavior. Recently, we demonstrated that the value-enhancing effects of nicotine are well characterized by reinforcer demand models, and that the value-enhancing effects of the smoking cessation aid bupropion (Zyban®) are distinct from those of nicotine and differ between the sexes (Barrett et al., 2017). The present study evaluated potential sex differences in the enhancement effects of nicotine and varenicline (Chantix®) using a reinforcer demand methodology. The role of α4β2* and α7 nicotinic acetylcholine receptors (nAChRs) in the enhancing effects of nicotine and varenicline are also evaluated. Male and female rats (n=12/sex) were trained to lever-press maintained by sensory reinforcement via visual stimulus (VS) presentations. Changes in VS value following nicotine and varenicline administration were assessed using an established reinforcer demand approach. Subsequently, the effects of antagonism of α4β2* and α7 nAChRs on varenicline and nicotine induced enhancement active lever-pressing was assessed using a progressive ratio schedule. Nicotine and varenicline enhanced VS demand equivalently between the sexes as evaluated by reinforcer demand. However, α4β2* receptor antagonism attenuated value enhancement by nicotine and varenicline in females, but only of nicotine in males.

Aims: Extracts of Chelidonium majus , extensively used in traditional systems of medicine, have been tested for their possible anti-tumor, hepato-protective and anti-genotoxic effects, to investigate their potential in cancer therapy. Although the highly active antiretroviral therapy (HAART) has changed the natural history of HIV infection, several adverse events may limit its efficacy. Antiretroviral drugs are associated with increased risk of severe hepatotoxicity. These complications increase morbidity and mortality significantly. Current study evaluated the effect of Chelidonium majus on metabolic alterations induced in mice subjected to HAART. Methodology: Four-weeks old male Swiss Webster mice, weighing approximately 28-30 g, provided by the Central Animal Laboratory of the State University of Maringa, were used in the experiments. The drug in the form of mother tincture was prepared with the juice of the root of C. majus , mixed in equal parts of grain alcohol (P.A.) and comprised 12 animals per experimental group: (I) animals treated with HAART diluted in 1.2 mL water gavage/day, (II) animals treated with HAART diluted in 1.2 mL water gavage/day + C. majus diluted 1x10 12 in water 1.0 mL, daily added

OBJECTIVE To determine the relationship between reduced retinal autofluorescence and the dense scotomas in Stargardt disease. METHOD Retrospective review of patients with Stargardt disease seen between 2000 and 2004 at a tertiary referral center. PATIENTS All 32 patients with Stargardt who underwent both autofluorescence (AF) imaging and scotoma mapping were included. RESULTS Forty-seven percent of eyes had dense scotomas corresponding to the areas of loss of AF (27% corresponding to the area of uniform loss of AF, and 20% corresponding to the area of uniform plus mottled loss of AF). Fifty-three percent of eyes did not have a good correlation of the dense scotoma with the area of loss of AF. Both eyes were judged to be in the same group in 75% of patients. CONCLUSIONS Areas of reduced AF are measurable, but are not equivalent to the area of dense scotoma in 53% of patients. Measures of retinal sensitivity and scotomas are necessary for defining retinal functional involvement in Stargardt disease.

PurposeQuantification of tissue stiffness may facilitate identification of abnormalities in orofacial muscle tone and thus contribute to differential diagnosis of dysarthria. Tissue stiffness is affected by muscle tone as well as age-related changes in muscle and connective tissue.MethodThe Myoton-3 measured tissue stiffness in 40 healthy adults, including equal numbers of men and women in each of two age groups: 18–40 years and 60+ years. Data were collected from relaxed muscles at the masseter, cheek, and lateral tongue surfaces in two positions: reclined on the side and seated with head tilted.ResultsTissue stiffness differed across age, sex, and measurement site with multiple interaction effects. Overall, older subjects exhibited higher stiffness coefficients and oscillation frequency measures than younger subjects whereas sex differences varied by tissue site. Effects of body position were inconsistent across tissue site and measurement.ConclusionsAlthough older subjects were expected to have lower muscle tone, age-related nonmuscular tissue changes may have contributed to yield a net effect of higher stiffness. These data raise several considerations for the development of accurate normative data and for future diagnostic applications of tissue stiffness assessment.

To determine if the severity of diabetic macular edema influences the effectiveness of subthreshold micropulse (STMP) laser treatment.A total of 63 eyes of 58 patients with diabetic macular edema were divided into two groups based on their initial central foveal thickness (CFT). Group 1 had CFT ≤400 μm, group 2 had CFT400 μm. The change from baseline in CFT and visual acuity were compared at 3, 6 and 12 months follow-up. Patients were considered for retreatment with micropulse laser at 3 months if macular edema had not improved. Patients were considered for rescue anti-VEGF injections if there was clinically significant macular edema at 6 months follow-up. Number of laser retreatments, injections, and any adverse effects from STMP laser were recorded.Group 1 (n=33) experienced an average of 55 μm reduction in CFT and 0.2 log MAR gain in visual acuity at 12 months (P0.001). No patient required rescue anti-VEGF injections. Group 2 (n=30) experienced no significant change in CFT or visual acuity by 6 months despite retreatment with STMP in 19 eyes. From 6 to 12 months follow-up, all the patients in group 2 received rescue Bevacizumab injections that resulted in 307 μm reduction in CFT and 0.3 log MAR improvement in visual acuity (P0.001). No adverse effects from STMP laser were recorded.Severity of edema can influence the effects of STMP laser. STMP monotherapy is safe and effective in treating edema of mild to moderate severity.

To evaluate the visual outcomes and macular thickness change in patients with symptomatic chronic central serous chorioretinopathy after treatment with a subthreshold MicroPulse diode laser.In this retrospective, interventional case series, 10 patients were treated with the subthreshold 810-nm diode MicroPulse laser. Selected patients had symptomatic disease that may or may not have involved the foveal center. The MicroPulse laser was applied to the areas of leakage seen on fluorescein angiogram, over the areas of clinical neurosensory detachment, and/or pigment epithelial detachments. Pretreatment and posttreatment vision, change in maximum macular thickness, number of treatment sessions, and number of laser spot applications were recorded. Patients were excluded if they did not attend follow-up, had other confounding macular diseases, were using steroid medications, or application of another treatment modality had been used (i.e., photodynamic therapy or anti-vascular endothelial growth factor medication).Ten patients met the inclusion criteria, with 1 patient treated in both eyes. Three patients were excluded for lack of follow-up, one for the use of systemic steroids, and one for treatment with anti-vascular endothelial growth factor injection. Maximum macular thickness decreased after subthreshold MicroPulse laser treatment between 20 μm and 338 μm (mean = 97 μm decrease, P = 0.0046) in 11 treated eyes.Subthreshold diode MicroPulse laser is a potential treatment option for patients with symptomatic chronic central serous chorioretinopathy.

[1] We report measurements of ablation rates of the bottom of two supraglacial lakes and of temperatures at different depths collected during the summers of 2010 and 2011 in west Greenland. To our knowledge, this is the first time that such data sets are reported and discussed in the literature. The measured ablation rates at the bottom of the two lakes are of the order of ∼6 cm/day, versus a rate of ∼2.5–3 cm/day in the case of bare ice of surrounding areas. Though our measurements suggest the presence of a vertical temperature gradient, it is not possible to draw final conclusions as the measured gradient is smaller than the accuracy of our temperature sensors. In-situ measurements are compared with the results of a thermodynamic model forced with the outputs of a regional climate model. In general, the model is able to satisfactorily reproduce the measured quantities with RMSE of the order of 3–4 cm for the ablation and ∼1.5°C in the case of water temperature. Our results confirm that the ablation at the bottom of supraglacial lakes plays an important role on the overall lake volume with the ablation in the case of ice covered by a lake being 110–135% of that over bare ice at nearby locations. Beside ice sheet hydrological implications, melting at the bottom of a supraglacial lake might affect estimates of lake volume from spaceborne visible and near-infrared measurements.

Plus de 70% des patients cancereux presentent des douleurs lorsque la maladie est avancee. La majorite de ces patients ont plus de 60 ans. L'echelle de traitement proposee par l'Organisation mondiale de la Sante, diffusee en Suisse par la Ligue suisse contre le Cancer, permet — lorsque les medicaments, salicyles, codeine et morphine sont donnes a doses adaptees et a heures fixes — de controler plus de 90% des douleurs. En phase terminale, la dyspnee, l'agitation ou la confusion et l'angoisse posent de difficiles problemes aux medecins, mais actuellement des traitements simples permettent d'y pallier.

Phelan, P. D. and Steiner, N. (1976).Aust. paediat. J., 12, 72–76. Assessing obstructive airways disease in children. Which test is best? Tests for airways obstruction based on a forced expiration were carried out in 60 children with asthma and 15 with cystic fibrosis to determine which test or tests were the best in identifying children with airways disease, to determine the severity of disease and to document response to therapy. Maximum expiratory flow, measured from a maximum expiratory flow volume curve, and maximum mid-expiratory flow measured from a spirogram, were the most sensitive tests in detecting early disease. Maximum expiratory flow measured from the maximum expiratory flow volume curve, combined with measurement of lung volumes, gave the best overall estimate of severity. Peak expiratory flow rate measured with a Peak Flow Meter, because of its simplicity of measurement, was the most useful test for routine use in the clinic and consulting room to monitor severity and response to therapy.

Diphenylacety Idiethylaminoethanol-hydrochloride (T), a new synthetic drug, has been assayed on barium chloride, as well as by indirect stimulation through parasympathetic nerves (acetylcholine, acetylbetamethylcholine, pilocarpine, eserine, electric stimulation of the vagus). A continuous stimulation with rise of tonus and increased contractions of the hollow viscera was provided by the slow intravenous injection of small amounts of a.c. or eserine. This method proved to be most useful for testing the relaxing effects of T as well as of other drugs. Motility of the stomach, small and large intestine, gall bladder and urinary bladder were recorded. In the case of the gall bladder relatively small doses of T produced most marked relaxation. This latter observation was confirmed in experiments on normal unanesthetized dogs reported elsewhere, in which the normal gall bladder was visualized fluoroscopically and in which T relaxed the resting as well as the physiologically contracted gall bladder (5). A comparison of the M.L.D. and of effective relaxing doses of atropine and of T reveals that a relatively smaller dose of T is more effective in causing relaxation. T in doses found to be non-toxic in normal animals compared well with the effects of 1 mg. of atropine sulfate and 2.5 mg. of papaverine hydrochloride administered in a number of experiments (see Table I and III, and Fig. 2). The effect of T (10–20 mg.) on salivary secretion of the dog was found to be slight and transient, while 1 mg. of atropine sulfate abolished it completely. In trials on ourselves 150 mg. of T per os produced no perceptible dryness of the mouth.

Measurements of maximum expiratory flows and lung volumes were made on 6 occasions at weekly intervals in 14 children who were known over several years to have severe chronic asthma. 11 of the 14 had persistent lung hyperinflation and marked reduction in maximum expiratory flow rates although there was considerable variability in individual measurements. The other 3 usually had hyperinflation and reduced expiratory flow rates. 3 times daily measurements at home of peak expiratory flow rate did not contribute further to assessment. The results of the study indicated that one or two measurements of maximum expiratory flow calculated from a maximum expiratory flow volume curve and of lung volumes recorded in a body plethysmograph are of value in identifying the child with severe chronic asthma.

The calcium-dependent regulator protein, calmodulin, is a 17,000 molecular weight polypeptide which binds calcium and has been shown to confer calcium sensitivity on contractile and other proteins. In the present study, we have examined the presence and subcellular distribution of this protein in preparations of human platelets. Calmodulin was quantified using a two-stage phosphodiesterase assay. Whole platelets contained 1.33 +/- 0.06 units calmodulin per 10(6) platelets or 26.5 +/- 3.4 fg calmodulin per platelet. The distribution of calmodulin in the platelet was predominantly soluble with over 80 percent of calmodulin activity in the soluble fraction of the cell. There was no apparent difference in the distribution of calmodulin between soluble and particulate compartments in recalcified platelet homogenates compared to homogenates in EDTA. Indirect immunofluorescent studies with monospecific antisera to dinitrophenylated calmodulin showed intense staining of platelets in a diffuse pattern. The identification of calmodulin in platelets raises the possibility that this protein may participate in calcium-dependent reactions important in platelet aggregation and release.