Зондовая микроскопия в исследовании изменений роста, подвижности, метаболизма и секреции раковых клеток

Современные методы сканирующей зондовой микроскопии позволяют получить детальную картину топологии живых клеток, в том числе раковых клеток, с нанометровым пространственным разрешением в процессе их роста. Развитие методов высокоскоростной атомно-силовой микроскопии дало возможность получать изображение клеток с миллисекундным пространственным разрешением. Вместе с тем сканирующая капиллярная (ион-проводящая) микроскопия позволяет исследовать шероховатую поверхность живых клеток за счет изменения протекающего ионного тока, при этом практически исключая силовое воздействие на клетку. Использование сканирующей капиллярной микроскопии в исследовании раковых клеток открывает новые возможности для скрининга лекарств, для получения новых данных о влиянии изменения внешних условий на кинетику роста опухоли, данных о жизнедеятельности клеток. Modern methods of scanning probe microscopy make it possible to obtain a detailed pattern of the vital cells topology including cancer cells with a nanoscale spatial resolution during their growth. The development of high-speed atomic force microscopy enabled to produce images of cells with millisecond spatial resolution. Besides, it is possible to study a rough surface of vital cells by changing the ion current flow without force action on a cell using scanning capillary microscopy (ion-conducting microscopy). The use of scanning capillary microscopy in the study of cancer cells opens up new opportunities for drugs screening in order to obtain new data on the influence of external conditions changes on the kinetics of tumor growth and the new data on the vital activity of cells.

AB1088 NAIL PSORIASIS: THE UNDERESTIMATED DISORDER IN PSORIASIS AND PSORIASIS ARTHRITIS. CAN ULTRASOUND AND CAPILLARY MICROSCOPY IN PATIENTS WITH NAIL PSORIASIS SPEED UP OUR DIAGNOSIS AND THERAPY?

Background:Nail psoriasis is an extreme diagnostic and therapeutic challenge and represents an enormous physical and psychological burden for affected patients.50% of patients with psoriasis vulgaris develop nail involvement (NailPso)during the course of their disease. NailPso is the strongest predictor of psoriatic arthritis (PsA). Through the synovio-enthesial concept we have learned that there is an anatomical-pathophysiological relationship between DIP joint, extensor tendon and nail matrix. We have observed in daily practice that hypervascularization (HV) in ultrasound Power Doppler (US-PD)the nail matrix may be a pathognomonic element in its own right. There are no data on this in the literature.Objectives:Is there a difference in the ultrasound PD examination of the DIP joint and nail area and in the capillary microscopy of the corresponding nail fold in patients with psoriasis vulgaris and nail psoriasis versus patients with psoriasis vulgaris without nail psoriasis.Methods:Monocentric prospective study of all consecutive patients with psoriasis vulgaris who have come to a rheumatic practice to clarify a PsA. Inaddition to demographic data, assessments (PASI,DLQI,CASPAR,GEPARD, DAS28, SJ,TJ,FFBH), clinical examination, a standardized ultrasound PD examination and capillary microscopy of the affected fingertips in PsO patients suffering from nail psoriasis was performed as well as corresponding examinations of the 2nd and 3rd finger right in PsO patients without nail involvement.Results:79 patients could be included during the study period. Thereof 25 PsO patients without nail involvement and 44 PsO patients with nail involvement. Since the patients were examined consecutively, the difference results. There was no difference in age, BMI and sex in both groups (Pso and NailPso). The Caspar criteria as classification criteria for a PsA were positive in 65% of the NailPso patients and positive in 50% of all PsO patients without nail infestation. Hypervascularization in the US-PD examination in the area of the nail matrix could be seen significantly more frequently in NailPso compared to non-NagelPso patients. Such a difference did not exist in the HV of the extensor tendons.Capillary microscopy showed a significant difference in the number of torsions/twist capillaries in NailPso compared to Pso patients without NailPso.Hypervascularization of the nail matrix is seen significantly more frequently in patients with psoriasis of the nail than in patients without psoriasis of the nail. Such a difference does not exist in DIP joint -extensor tendon- enthesitis. At the same time, torsions are significantly more frequently seen in capillary microscopy in NailPso than in patients without NailPso.Conclusion:The US-PD examination is a simple and non-invasive procedure which can be performed routinely in daily practice. The hypervascularization of the nail matrix should also make one think of nail psoriasis in the early stage of PsO, in order to be able to start early an appropriate therapy for this very stigmatizing and therapeutically extremely difficult manifestation of PsO. It seems to occur independently of extensor tendon synovitis as an independent manifestation phenomenon.The occurrence of torsions in capillary microscopy >50 % also seems to be groundbreaking for a NagelPso, whereby capillary microscopy is a temporal challenge in daily routine.References:§ The present study (7734-BO-S2018 Ethics Commission of the MHH, Medical School Hannover, Germany) contains parts of the PhD thesis of M.TöllnerDisclosure of Interests:Becker-Capeller Detlef Grant/research support from: Novartis, Speakers bureau: Pfizer, Soham El-Nawab-Becker: None declared, Malo Toellner: None declared, Andreas Kleinheinz: None declared, Torsten Witte: None declared

THU0357 NAILFOLD CAPILLARY MICROSCOPY HAS LIMITED PROGNOSTIC VALUE IN PREDICTING FUTURE DEVELOPMENT OF CONNECTIVE TISSUE DISEASE IN CHILDREN WITH RAYNAUD’S PHENOMENON.

Background:For adults with Raynaud’s phenomenon (RP), nailfold capillary microscopy (NCM) is established to be an effective method for differentiating between PRP and SRP (1,2). Although Raynaud’s phenomenon (RP) is very common in childhood, studies on diagnostic methods to differentiate between primary RP (PRP) and secondary RP (SRP) at a young age are scarce (3,4).Objectives:The general aim of this study was to determine the prognostic value of nailfold capillary microscopy (NCM) in addition to antinuclear antibodies (ANAs) for later development of connective tissue diseases (CTD) in children with RP.Methods:This was a case-control study, in which 83 patients diagnosed with RP and having undergone NCM in childhood were retrospectively included. Based on whether they were diagnosed with a connective tissue disease (CTD) during follow-up, they were classified as PRP or SRP. PRP and SRP patients were compared on demographics, NCM and ANA positivity. Variables associated with SRP were included in a multivariate logistic regression model. Predictive values were calculated for NCM, ANA positivity and the combination of NCM and ANA positivity.Results:At the time of the baseline NCM, the mean age of the RP patients was 15.4±2.3 years. Averagely 6.4±3.2 years after the baseline NCM, 65 of the 83 patients were classified as PRP and 18 as SRP. The most common CTDs were MCTD and undifferentiated CTD. ANA positivity was associated with SRP (p<0.001). Of the NCM parameters, only capillary loss was associated with SRP (p=0.01). Abnormal numbers of dilated capillaries, giant capillaries and haemorrhages were not significantly associated with SRP. In a multivariate logistic regression model, only ANA positivity was predictive for SRP (OR 11.19, CI 3.07-40.79). ANA alone had a sensitivity of 66.7% and a specificity of 85.9% for SRP. ANA combined with capillary loss had a sensitivity of 33.3% and a specificity of 96.8%.Conclusion:This study demonstrates that childhood RP is primary in most cases. Whereas RP in adulthood is most strongly associated with SSc, children with RP seem to be at risk of developing other CTDs with less apparent NCM abnormalities. Dilated capillaries, giant capillaries and haemorrhages on NCM are not associated with the spectrum of CTDs that children are at risk for, and do not differentiate between primary and secondary RP. Although capillary loss on NCM is associated with SRP, capillary loss may add little to the predictive value of serology. To clarify which NCM parameters are helpful for early detection of SSc-like CTDs, additional research is required.References:[1]Koenig M, Joyal F, Fritzler MJ, Roussin A, Abrahamowicz M, Boire G, et al. Autoantibodies and microvascular damage are independent predictive factors for the progression of Raynaud’s phenomenon to systemic sclerosis: A twenty-year prospective study of 586 patients, with validation of proposed criteria for early systemic sclerosis. Arthritis Rheum. 2008;58(12):3902–12.[2]Pavlov-Dolijanovic S, Damjanov NS, Stojanovic RM, Vujasinovic Stupar NZ, Stanisavljevic DM. Scleroderma pattern of nailfold capillary changes as predictive value for the development of a connective tissue disease: a follow-up study of 3,029 patients with primary Raynaud’s phenomenon. Rheumatol Int. 2012;32(10):3039–45.[3]Jones GT, Herrick AL, Woodham SE, Baildam EM, Macfarlane GJ, Silman AJ. Occurrence of Raynaud’s phenomenon in children ages 12-15 years: Prevalence and association with other common symptoms. Arthritis Rheum. 2003;48(12):3518–21.[4]Pavlov-Dolijanović S, Damjanov N, Ostojić P, Sušić G, Stojanović R, Gacić D, et al. The prognostic value of nailfold capillary changes for the development of connective tissue disease in children and adolescents with primary Raynaud phenomenon: A follow-up study of 250 patients. Pediatr Dermatol. 2006;23(5):437–42.Disclosure of Interests:None declared

SAT0325 STELLATE BLOCKADE COMBINED TO ILOPROST AS SUPPORTIVE TREATMENT OPTION IMPROVES PAIN AND ISCHAEMIC SYMPTOMS IN PATIENTS WITH SYSTEMIC SCLEROSIS

Background:Peripheral ischaemia is a common symptom in systemic sclerosis (SSc) patients with risk of development of digital ulcers (DU). For its treatment, intravenous iloprost is the most effective option. Accompanying pain symptoms worsen the ischaemic symptoms, so a combination with anaesthetic procedures may improve ischaemic status and the subjective sensation of raynaud and pain. The aim of this study was to observe the impact of a combined treatment of iloprost with stellate blockade (ILOST) in improvement of ischaemic symptoms compared to iloprost treatment only (ILO).Objectives:To evaluate efficacy of the ILOST treatment on changes in vascularisation and sensation of patients with SSc and indication for vasodilatative treatment with Iloprost.Methods:Twenty SSc-patients with indication for ILO-treatment (prophylactic or due to digital ulcerations (DU)) will be included in a prospective observational study. Patients will be offered to combine ILO with stellate blockade (ILOST). Beside documentation of disease activity characteristics (mRSS, number of DU, capillary microscopy at baseline, after ILO-treatment and at week 12), patients are assessed using fluorescence-optical imaging (FOI) as innovative method for illustration of changes in microvascularisation and patient reported outcomes (DASH, VAS) at week 12.Results:This interims analysis includes the result of the first 11 patients treated. Mean baseline characteristics (age and gender) are well balanced. Iloprost treatment was initiated due to prophylactic treatment to avoid DU in all patients. 100% of the patients in the ILOST-group were diagnosed as limited SSc compared to 60% in the ILO-group (diffuse type with 40%). All patients showed abnormalities in capillary microscopy (ILOST group: 83,3% late pattern 16.7% active pattern; ILO group: 80% late pattern, 20% early pattern). MRSS was low in both groups with 1.8, the disease duration in mean 15.3 years in the ILOST-group compared to 13.2 in the ILO-group, respectively. In both groups, no new DU occurred in the 12-week follow-up. Improvement in VAS pain was reported in 83% of the patients in the ILOST group compared to 60% in the ILO group. DASH improved with a mean of 5.5 points in the ILOST group compared to 3 points in the ILO group. FOI was compared individual at both arms in the ILOST group only. The arm with stellatum blockade showed a pronounced increase of FOI signals of 5% in mean whereas the opposite site showed a decrease of the signal shortly after ILO treatment indicating a pronounced increase of vascularisation in the ILOST treated body site.Conclusion:A new treatment approach to improve acute ischaemic symptoms was tested by combining stellate blockade to iloprost treatment. No new DU occurred up to 12 weeks after treatment in all patients of both groups indicating the relevance of iloprost as effective vascular dilatative therapy in SSc. The additional intervention was well tolerated and asked to repeat. Subjective sensation on pain of the hands as well as DASH was improved in the combined group. FOI showed a relevant increase in vascularisation in the blockade arm compared to the opposite site in which signals decreased indicating a stronger effect of the combined treatment for improvement of vascularisation.Disclosure of Interests:Franziska Höcketstaller Grant/research support from: Rheumazentrum Rhein-Main, Ulf Henkemeier: None declared, Michael Zimmermann: None declared, Harald Burkhardt Grant/research support from: Pfizer, Roche, Abbvie, Consultant of: Sanofi, Pfizer, Roche, Abbvie, Boehringer Ingelheim, UCB, Eli Lilly, Chugai, Bristol Myer Scripps, Janssen, and Novartis, Speakers bureau: Sanofi, Pfizer, Roche, Abbvie, Boehringer Ingelheim, UCB, Eli Lilly, Chugai, Bristol Myer Scripps, Janssen, and Novartis, Frank Behrens Grant/research support from: Pfizer, Janssen, Chugai, Celgene, Lilly and Roche, Consultant of: Pfizer, AbbVie, Sanofi, Lilly, Novartis, Genzyme, Boehringer, Janssen, MSD, Celgene, Roche and Chugai, Ulrich Drott: None declared, Michaela Köhm Grant/research support from: Pfizer, Janssen, BMS, LEO, Consultant of: BMS, Pfizer, Speakers bureau: Pfizer, BMS, Janssen, Novartis

Prevalence and diagnostic value of nail fold capillary microscopy in hereditary hemorrhagic telangiectasia: A retrospective study

Abnormal vasculature is a key feature of hereditary hemorrhagic telangiectasia (HHT) and can also present in the nail fold capillary beds. However, the exact prevalence and the clinical diagnostic value in HHT are still largely unknown. The nail fold can be easily and noninvasively inspected with a capillary microscope. We therefore retrospectively assessed the prevalence and diagnostic value of abnormal nail fold capillaries in all patients who were screened between January 2000 and July 2017 for the presence of HHT and underwent capillary microscopy in St Antonius Hospital, The Netherlands. Capillary microscopy results and clinical characteristics were extracted from medical files and the prevalence of abnormal nail fold capillaries was calculated and the diagnostic value of the Curaçao criteria with and without capillary microscopy results was assessed. Of the 1761 individuals screened, 923 (52%) were diagnosed with a clinical and/or genetic HHT diagnosis. In these patients, capillary microscopy was normal in 23% ( n = 218), enlarged loops were seen in 11% ( n = 99), and giant loops in 66% ( n = 606). The sensitivity and specificity of the Curaçao criteria for the diagnosis of HHT without capillary microscopy results were 96% and 90%, respectively. The addition of the presence of giant loops to the Curaçao criteria led to a small increase in sensitivity to 97% without affecting the specificity. In conclusion, the prevalence of nail fold abnormalities in patients with HHT is high. Capillary microscopy can be a useful, easy, and noninvasive diagnostic tool in HHT.

Применение сканирующей зондовой и капиллярной микроскопии в международном сотрудничестве

The scientific collaboration of the probe microscopy group of Lomonosov Moscow State University and the scientific group from the Sharif University of Technology (Tehran) in the framework of the Russian-Iranian project was extremely fruitful. For three years of the project the original results on local surface modification in thin films due to the use of scanning capillary microscopy were obtained. Also, a device for combined probe and capillary microscopy has been developed [1, 2].