Next-generation sequencing (NGS) of cell-free circulating tumor DNA and tumor tissue in patients with advanced urothelial cancer: a pilot assessment of concordance.

BACKGROUND: Advances in cancer genome sequencing have led to the development of various next-generation sequencing (NGS) platforms. There is paucity of data regarding concordance of different NGS tests carried out in the same patient. METHODS: Here, we report a pilot analysis of 22 patients with metastatic urinary tract cancer and available NGS data from paired tumor tissue [FoundationOne (F1)] and cell-free circulating tumor DNA (ctDNA) [Guardant360 (G360)]. RESULTS: The median time between the diagnosis of stage IV disease and the first genomic test was 23.5 days (0-767), after a median number of 0 (0-3) prior systemic lines of treatment of advanced disease. Most frequent genomic alterations (GA) were found in the genes TP53 (50.0%), TERT promoter (36.3%); ARID1 (29.5%); FGFR2/3 (20.5%), PIK3CA (20.5%) and ERBB2 (18.2%). While we identified GA in both tests, the overall concordance between the two platforms was only 16.4% (0%-50%), and 17.1% (0%-50%) for those patients (n = 6) with both tests conducted around the same time (median difference = 36 days). On the contrary, in the subgroup of patients (n = 5) with repeated NGS in ctDNA after a median of 1 systemic therapy between the two tests, average concordance was 55.5% (12.1%-100.0%). Tumor tissue mutational burden was significantly associated with number of GA in G360 report (P < 0.001), number of known GA (P = 0.009) and number of variants of unknown significance (VUS) in F1 report (P < 0.001), and with total number of GA (non-VUS and VUS) in F1 report (P < 0.001). CONCLUSIONS: This study suggests a significant discordance between clinically available NGS panels in advanced urothelial cancer, even when collected around the same time. There is a need for better understanding of these two possibly complementary NGS platforms for better integration into clinical practice.

[Efficacy of walking training in patients with obliterating atherosclerosis of lower limb arteries]. / Éffektivnost' trenirovochnoi khod'by pri obliteriruiushchem ateroskleroze arterii nizhnikh konechnostei.

The authors assessed efficacy of treating patients with atherosclerosis obliterans of lower-limb arteries using the method of walking training. Our prospective study included a total of 84 patients suffering from intermittent claudication. They were divided in two groups. The Study Group (Group I) comprised 64 patients undergoing comprehensive treatment including walking training (WT) and the Control Group (Group II) consisted of 20 patients treated without a physical training programme. The patients were followed up monthly during three months, with registration of the pain-free walking distance (PFWD), maximum walking distance (MWD), and the ankle-brachial index (ABI). There were no baseline statistically significant differences between Group I and Group II patients in the parameters examined. The Study Group patients were found to have a considerable increase in the PFWD and MWD values, with the statistical significance of differences from the Control Group patients observed as early as at one month of treatment. The baseline PFWD value in the Study Group amounted to 132.4±14.7 m, being at 3 months 259.7±29.6 m (p<0.05); the MWD value in the Study Group initially equalled 170.2±12.3 m, amounting at 3 months to 364.6±39.2 m (p<0.05). The Study Group patients by the end of follow up were also found to have an increase in the ankle-brachial index by nearly 25% from the baseline level. The carried out treatment resulted in an increase in the number of patients with less severe stages of arterial insufficiency: initially stage IIa was diagnosed in 18 (28.2%) patients and at 3 months in 47 (73.4%) patients, with stage IIb being diagnosed in 46 (71.8 %) and 17 (26.6%) subjects, respectively. A conclusion was drawn that training walking considerably improved clinical manifestations of the disease. Taking into consideration availability of this therapeutic method, it should be recommended for patients suffering from intermittent claudication in long-term courses and in all cases where there are no contraindications thereto.

[Peculiarities of atherosclerotic lesions of lower limb arteries in patients with type 2 diabetes mellitus]. / Osobennosti ateroskleroticheskogo porazheniia arterii nizhnikh konechnostei u bol'nykh sakharnym diabetom 2 tipa.

The authors carried out a comparative study of the arterial vascular wall in patients presenting with atherosclerosis obliterans of lower-limb arteries on the background of diabetes mellitus and without it, thoroughly describing the methodology and reporting the results of the study. The obtained findings make it possible to increase the informative value of diagnostic measures and to more exactly determine therapeutic policy.

[Comparative assessment of antiaggregant efficacy of acetylsalicylic acid and clopidogrel in peripheral atherosclerosis].

Presented in the article are the results of studying antiaggregate activity of acetylsalicylic acid and clopidogrel in a total of 36 patients suffering from various-severity chronic arterial insufficiency of lower limbs on the background of atherosclerosis obliterans. The study was conducted prior to treatment for correct selection of a particular antiplatelet agent. The obtained results showed that clopidogrel was not always more efficient than acetylsalicylic acid, since there is individual sensitivity of each patients to a particular antiplatelet agent. Therefore, an individual approach is necessary to conservative therapy of arterial insufficiency of lower limbs. For some patients it is preferable to administer clopidogrel, for others - acetylsalicylic acid. In a series of cases combined treatment is justified, while some patients having low sensitivity to both acetylsalicylic acid and clopidogrel require careful selection of alternative agents influencing other thrombocyte receptors.

[Detection of genetic determinants that define the difference of near-isogenic Triticum aestivum L. Lines in photoperiodic sensitivity].

Identification of genetic determinants that define different degrees of line sensitivity to the photoperiod was conducted on material of near-isogenic lines of the soft hexaploid wheat Triticum aestivum L. using SSR markers and markers specific to the Vrn and Ppd genes. It was established that the Ppd-s line contains a dominant Ppd-Dla allele located on chromosome 2D. This allele is characterized by a vast deletion in the gene promoter region. For two other lines (Ppd-m and Ppd-w), introgression of the Ppd-B1 gene on chromosome 2B was shown from the parental Sonora variety, which is slightly sensitive to the length of the day; however, the previously described Ppd-Bla. 1 allele was not found. Another polymorphism that can cause weak photoperiodic sensitivity, an increased amount of the Ppd-B1 gene copies, was detected for these lines.

[Role of low-dose acetylsalicylic acid in comprehensive postoperative treatment of patients with lower-limb chronic arterial insufficiency].

The study comprised a total of 107 patients (all men) after endured femoropopliteal bypass grafting above the genicular fissure with a synthetic stent graft manufactured by the Gore Company for stage IIB and III chronic arterial insufficiency of the lower extremities according to the Fontain-Pokrovsky classification. Group One I (control group) was composed of fifty-four patients permanently taking in the postoperative period at the out-patient stage pentoxiphylline (trental 400 mg 1 tablet 3 times daily) and xantinol nicotinate at a dose of 150 mg one tablet thrice daily. Group Two (Study Group) consisted of fifty-three patients taking after reconstructive vascular surgery at the out-patient stage in addition to pentoxiphylline and xantinol nicotinate acetylsalicylic acid (cardiomagnil 75 mg 1 tablet once daily). The Control Group patients within 3 to 6 months of follow up were found to have a considerable progressing improvement of the functional abilities of the microcirculatory bed requiring in 44 (81.5%) cases hospitalization to the Surgical Department for intensive vascular therapy. Despite this fact four (7.4%) patients within the time frame from 6 to 9 months after surgery developed thrombosis of the vascular implant requiring a repeat surgical intervention. In the Study Group patients, the degree of functional capabilities of the microcirculatory bed in the postoperative period was less considerable, reaching the maximum after 10-12 months of follow up, with eighteen (34.0%) patients requiring hospitalization for additional vascular therapy to perform. There were no cases of implants' thrombosis in the Study Group patients. Pathological alterations in the functional state of the peripheral vascular bed correlated with viscosimetric indices and activity of blood platelet aggregation. The addition of antithrombocytic agents to conservative postoperative therapy considerably improved the outcomes of surgical treatment.

Beta cell-specific Znt8 deletion in mice causes marked defects in insulin processing, crystallisation and secretion.

AIMS/HYPOTHESIS: Zinc is highly concentrated in pancreatic beta cells, is critical for normal insulin storage and may regulate glucagon secretion from alpha cells. Zinc transport family member 8 (ZnT8) is a zinc efflux transporter that is highly abundant in beta cells. Polymorphisms of ZnT8 (also known as SLC30A8) gene in man are associated with increased risk of type 2 diabetes. While global Znt8 knockout (Znt8KO) mice have been characterised, ZnT8 is also present in other islet cell types and extra-pancreatic tissues. Therefore, it is important to find ways of understanding the role of ZnT8 in beta and alpha cells without the difficulties caused by the confounding effects of ZnT8 in these other tissues. METHODS: We generated mice with beta cell-specific (Znt8BKO) and alpha cell-specific (Znt8AKO) knockout of Znt8, and performed in vivo and in vitro characterisation of the phenotypes to determine the functional and anatomical impact of ZnT8 in these cells. Thus we assessed zinc accumulation, insulin granule morphology, insulin biosynthesis and secretion, and glucose homeostasis. RESULTS: Znt8BKO mice are glucose-intolerant, have reduced beta cell zinc accumulation and atypical insulin granules. They also display reduced first-phase glucose-stimulated insulin secretion, reduced insulin processing enzyme transcripts and increased proinsulin levels. In contrast, Znt8AKO mice show no evident abnormalities in plasma glucagon and glucose homeostasis. CONCLUSIONS/INTERPRETATION: This is the first report of specific beta and alpha cell deletion of Znt8. Our data indicate that while, under the conditions studied, ZnT8 is absolutely essential for proper beta cell function, it is largely dispensable for alpha cell function.

[Regional venous hypertension as a factor of compensation for microcirculatory impairments in chronic obliterating diseases of lower-limb arteries (state of the art)].

A thorough analysis of the literature and own data confirmed a lively interest of vascularsurgeons in alterations in venous haemodynamics in patients presenting with chronic obliterating diseases oflower-limb arteries. The presence of close relationships between impaired arterial and venous blood flow and a compensatory pattern of the development of venous hypertension in the ischaemia zone was proved. This should be taken into consideration during conservative therapy of patients sufferingfrom chronic obliterating diseases of lower-limb arteries, and while working out novel types of surgical interventions, as well as for more adequate interpretation of pathophysiological peculiarities of the existing disease.

[Prediction of remote results after surgical management of patients presenting with atherosclerosis obliterans of the lower-limb arteries].

Forecasting the development of implants' narrowing appears to be one of the most important problems encountered in vascular surgery today. The present study was therefore undertaken to work out a comprehensive diagnostic methodology making it possible to reliably predict the development of stenosis of vascular anastomoses during the postoperative period in patients presenting with atherosclerosis obliterans. The work was based upon the findings obtained by examining a total of seventy-nine patients diagnosed with atherosclerosis obliterans of the lower-limbs arteries, also including assessment of a large number of haemodynamic and immunological parameters both before and after reconstructive vascular interventions. The study showed substantial dynamics of a series of immunological and haemodynamic indices on the background of the operation and during the postoperative period. Based on the mathematical processing of the obtained data, the authors managed to reveal the factors playing a leading role in the development of stenosis, having also worked out a prognostic algorithm making it possible with high reliability to predict the development of stenosis of arterial anastomoses.

[Peculiarities of peripheral haemodynamics in patients with varicose disease of lower-limb veins complicated by trophic ulcers].

The work was dedicated to studying local, regional and systemic circulatory disorders in patients suffering from varicose disease of the lower extremities veins with accompanying trophic ulcers. The possibility of performing this work was determined by a diagnostic complex possessing high informative value (especially it concerns studying microcirculation). Based on the obtained findings we managed not only to evaluate severity of microcirculatory disorders but also to characterize them qualitatively, in particular to reveal the so-called blood microshunting phenomenon. Pathophysiologically important is the development of inflammatory hyperaemia in the area of trophic ulcers, which is prognostically favourable for their healing. Proceeding from the above-mentioned, utterly true turned out the conclusion concerning the necessity to include microcirculation-improving pharmaceutical substances into the program of treatment of patients suffering from varicose disease of the lower-limb veins, especially those presenting with concomitant trophic ulcers, along with surgical intervention.

alpha-lipoic acid regulates AMP-activated protein kinase and inhibits insulin secretion from beta cells.

AIMS/HYPOTHESIS: The antioxidant compound alpha-lipoic acid (alpha-LA) possesses antidiabetic and anti-obesity properties. In the hypothalamus, alpha-LA suppresses appetite and prevents obesity by inhibiting AMP-activated protein kinase (AMPK). Given the therapeutic potential of alpha-LA for the treatment of type 2 diabetes and obesity, and the importance of AMPK in beta cells, we examined the effect of alpha-LA on pancreatic beta cell function. MATERIALS AND METHODS: Isolated rat islets and MIN6 beta cells were treated acutely (15-90 min) or chronically (18-24 h) with alpha-LA or the known AMPK-activating compounds 5'-amino-imidazole-4-carboxamide ribonucleoside (AICAR) and metformin. Insulin secretion, the AMPK-signalling pathway, mitochondrial function and cell growth were assessed. RESULTS: Acute or chronic treatment of islets and MIN6 cells with alpha-LA led to dose-dependent rises in phosphorylation of the AMPK alpha-subunit and acetyl CoA carboxylase. Chronic exposure to alpha-LA, AICAR or metformin caused a reduction in insulin secretion. alpha-LA inhibited the p70 s6 kinase translational control pathway, and inhibited MIN6 growth in a manner similar to rapamycin. Unlike AICAR and metformin, alpha-LA also acutely inhibited insulin secretion. Examination of the effect of alpha-LA on mitochondrial function showed that acute treatment with this compound elevated reactive oxygen species (ROS) production and enhanced mitochondrial depolarisation induced by Ca(2+). CONCLUSIONS/INTERPRETATION: This study is the first to demonstrate that alpha-LA directly affects beta cell function. The chronic effects of alpha-LA include AMPK activation and reductions in insulin secretion and content, and cell growth. Acutely, alpha-LA also inhibits insulin secretion, an effect probably involving the ROS-induced impairment of mitochondrial function.

[Antioxidant and metabolic therapy of lower limb chronic obliterative arterial disease].

Metabolic therapy is one of several therapeutic strategies that can be used in patients with lower limb chronic obliterative arterial disease (COAD). Tanakan (EGb76) has demonstrated high efficiency and safety in 9 controlled clinical trials and author's clinical series in patients with lower limb COAD. Improvement with significant increase of pain-free walking distance and beneficial safety profile was observed in all cases. Tanakan (EGb76) is highly effective conventional therapeutic agent for memory and attention disorders, dizziness and balance disturbances in adults and elderly patients. Given a common concomitance of these disorders and lower limb COAD, Tanakan can be recommended as a drug of choice for treatment of chronic obliterative arteriopathy.

[Functional flowmetry in determining the indications for reconstructive surgical interventions on the major arteries of the pelvis and lower limbs].

The work was based on the findings of the examination and surgical management of 69 patients with obliterating atherosclerosis of the lower limbs with various degree of chronic arterial insufficiency. The authors analyzed the main causes promoting development of early reocclusion of the bypasses and prostheses following reconstructive surgical operations. They also proved the role of high peripheral vascular resistance and, in particular, diminished functional possibilities of the microcirculatory-bed vessels in the genesis of early reocclusions in patients with chronic arterial insufficiency of the lower limbs. They also suggested a new technique of determining the carrying capacity of the microcirculatory channel, the use of which in the preoperative period makes it possible to predict development of early thrombotic complications, and in some cases to wisely abstain from carrying out a reconstructive vascular intervention due to high risk of its poor outcomes. It is preferable for such patients to undergo conservative pathogenetic preoperative therapy aimed at improving the functional possibilities of the microcirculatory vessels.

[Preoperative care of microcirculatory vessels in patients with lower limb arteriosclerosis obliterans].

The paper analyses diagnosis and treatment outcomes for 20 men aged from 55 to 76 with lower limb arteriosclerosis obliterans. Based on exhaustive laboratory and instrumental diagnostic data the authors conclude that 10-days course of Vazaprostan (20 mkg per day) for patients with IIb stage of chronic lower limb arterial insufficiency yields not only improvement in tissue metabolism, but a reduction of peripheral vascular resistance as well. Thus such therapy can be used for preoperative care before reconstructive vascular interventions in order to reduce the risk of implant thrombosis in early postoperative period.

[Conservative treatment of lower limb chronic obliterative atherosclerosis: strategy and tactics].

The paper presents approaches and purposes of conservative treatment of lower limb chronic arterial obliterative lesions. Special attention is given to antiplatelet agents. It is stressed that outpatient management comprises the most important part of this program.

[The use of pentoxyphylline in the treatment of patients with chronic obliterative diseases of lower limb arteries].

The review concerns the assessment of the efficacy of pentoxyphylline which is most popular today in the treatment of patients with chronic arterial lower limb insufficiency. The new data are presented on its action mode as are multiple examples of the use of pentoxyphylline in the treatment of patients with peripheral angiopathies.

Arteriovenous anastomoses of major crural vessels in humans.

Arteriovenous anastomoses of the major crural vessels were studied postmortem. The revealed anastomoses were examined by histological technique.

[Critical ischemia resulted from inadequate outpatient management of lower limb chronic arterial obliterations]. / Kriticheskaia ishemiia kak sledstvie neadekvatnogo lecheniia bol'nykh khronicheskimi obliteriruiushchimi zabolevaniiami arterii nizhnikh konechnostei na ambulatornom étape.

Management of critical and subcritical lower limb ischemia poses a challenge due to poor effectiveness and high costs of therapy. Problem solving supposes not only the development of effective surgical and medical management of severe arterial insufficiency, but also preventive efforts on early outpatient stage. The paper reviews 611 patients with lower limb arterial obliterative lesions and correlates age, sex, concomitant diseases, stage of arterial insufficiency and risk factors. Consideration of these parameters can be useful in choosing the most effective management strategy. Authors mention that in community outpatient facilities treatment effectiveness of chronic lower limb arterial obliterative lesions does not exceed 40%, white the success of conservative treatment according to the protocol developed in their clinics reaches 90%. It means that there is a real perspective of lowering the incidence of severe lower limb arterial insufficiency managed in hospital environment by improving preventive and therapeutic strategies at outpatient care level.