Ejaculation-sparing enucleation of benign prostate hyperplasia: is it almost feasible?

Introduction. Patients often request maintenance of antegrade ejaculation, and try to find the clinic, surgeon, and treatment to store it. Despite an established technique, the long-term results of ejaculation-sparing operations at a large prostate volume remain unstable. This is not allowed to give patients a guarantee in maintaining this component in their sexual life.Objective. To evaluate the frequency of ejaculation maintenance after laser ejaculatory-sparing prostate enucleation.Materials & methods. Since 2017 ejaculation-sparing laser enucleations have been performed in prostate volume (V pr > 80 cc) in 84 patients. Fifty-four patients (64%) had three lobes benign prostate hyperplasia (BPH) and two lobes — 30 patients (36%). Preoperative voiding parameters were IPSS score — 21.0 ± 2.7, QoL — 4.8 ± 0.6, residual urine volume (V res) — 139 ± 43 mL, Q max — 8.1 ± 2.0 mL. We estimate the result as positive if antegrade ejaculation remained after intervention. If ejaculation volume decreased, then as partially positive and negative — ejaculation was absent postoperatively. We conducted questionnaire and follow-up examination 3 – 6 months after intervention.Results. Thulium-fiber laser enucleations have been performed in all cases without any serious complications. Followup 3 – 6 months after operation showed V pr — 29 ± 4 cc, V res — 19 ± 17 mL, Q max — 19.1 ± 3.1 mL/sec. IPSS score — 8.1 ± 1.9 and QoL — 2.6 ± 0.7. There are the following technical remarks: 1) tissue-sparing in the verumontanum; 2) no or minor mechanical tissue tension during enucleation; 3) avoid additional resection in the bladder neck; 4) refuse total coagulation; 5) no urethral catheter tension after operation; 6) catheter balloon inflow only in the bladder, not in the fossa. Antegrade ejaculation has stored at 17 (20%) patients, partial ejaculation — at 34 (40%) patients, so overall efficacy is 60%. The presence of a median lobe is shown to be negative prognostic factor due to the absence of full antegrade ejaculation in all cases. A partial ejaculation was achieved at 21 patients. Based on the obtained results we have optimized the three-lobe prostate enucleation technique. Four from our initial 7 patients had full storage of ejaculation and 3 — partial.Conclusion. The possibility of BPH laser surgery to store ejaculation function at the request of the patients is modest. The presence of the median lobe makes the full ejaculation-sparing impossible. In cases of two lobes BPH full storage of the ejaculation was at 57% and partial at 43%, respectively. The following evaluation of the described technique is mandatory

Immunogenicity of recombinant fragment of orthopoxvirus p35 protein in mice

Despite the elimination of smallpox, orthopoxviruses continue to be a source of biological danger for humans, as cowpox and monkey pox viruses circulate in nature and the last virus can cause both sporadic cases of human diseases and outbreaks of smallpox-like infection. In addition, periodic vaccination is necessary for representatives of some professions (scientists studying pathogenic orthopoxviruses, medical personnel, etc.). Vaccination against smallpox virus with live vaccinia virus, which was widely used during the elimination of smallpox, induces the formation of long-term immunity in vaccinated people. However, providing a high level of protection, the vaccination is often accompanied by serious post-vaccination complications, the probability of which is particularly great for individuals with compromised immunity. In this regard, the development of preparations for the prevention and treatment of infections caused by orthopoxviruses remains important today. The aim of this study was to assess the immunogenicity in the mouse model of recombinant protein р35Δ12, designed previously on the base of the cowpox virus protein p35. It was previously shown that the protein р35Δ12 was recognized by fully human neutralizing anti-orthopoxviral antibody with high affinity. In this work, recombinant protein р35Δ12 produced in E. coli cells XL1-blue and purified by chromatography was used for two-time immunization of mice. Two weeks after the second immunization, blood samples were taken from mice and serum antibodies were analyzed. It was shown by ELISA and Western-blot analysis that immunized mice sera contained IgG antibodies specific to recombinant protein р35Δ12. Confocal microscopy showed that antibodies induced by the р35Δ12 protein were able to recognize Vero E6 cells infected with the LIVP-GFP vaccinia virus. In addition, the antibodies in the serum of immunized mice were able to neutralize the infectivity of the vaccinia virus LIVP-GFP in the plaque reduction neutralization test in vitro. These experiments have demonstrated promising properties of the р35Δ12 protein if it were used as a component of vaccine for prophylaxis of orthopoxvirus infections

Cystathionine beta-Synthase (CBS) Domain-containing Pyrophosphatase as a Target for Diadenosine Polyphosphates in Bacteria

Among numerous proteins containing pairs of regulatory cystathionine beta-synthase (CBS) domains, family II pyrophosphatases (CBS-PPases) are unique in that they generally contain an additional DRTGG domain between the CBS domains. Adenine nucleotides bind to the CBS domains in CBS-PPases in a positively cooperative manner, resulting in enzyme inhibition (AMP or ADP) or activation (ATP). Here we show that linear P-1,P-n-diadenosine 5&#39;-polyphosphates (Ap(n)As, where n is the number of phosphate residues) bind with nanomolar affinity to DRTGG domain-containing CBS-PPases of Desulfitobacterium hafniense, Clostridium novyi, and Clostridium perfringens and increase their activity up to 30-, 5-, and 7-fold, respectively. Ap(4)A, Ap(5)A, and Ap(6)A bound noncooperatively and with similarly high affinities to CBS-PPases, whereas Ap(3)A bound in a positively cooperative manner and with lower affinity, like mononucleotides. All Ap(n)As abolished kinetic cooperativity (non-Michaelian behavior) of CBS-PPases. The enthalpy change and binding stoichiometry, as determined by isothermal calorimetry, were similar to 10 kcal/mol nucleotide and 1 mol/mol enzyme dimer for Ap(4)A and Ap(5)A but 5.5 kcal/mol and 2 mol/mol for Ap(3)A, AMP, ADP, and ATP, suggesting different binding modes for the two nucleotide groups. In contrast, Eggerthella lenta and Moorella thermoacetica CBS-PPases, which contain noDRTGG domain, were not affected by Ap(n)As and showed no enthalpy change, indicating the importance of the DTRGG domain for Ap(n)A binding. These findings suggest that Ap(n)As can control CBS-PPase activity and hence affect pyrophosphate level and biosynthetic activity in bacteria.</p

Pre-steady-state kinetics and solvent isotope effects support the "billiard-type" transport mechanism in Na+-translocating pyrophosphatase

Membrane-bound pyrophosphatase (mPPase) found in microbes and plants is a membrane H+ pump that transports the H+ ion generated in coupled pyrophosphate hydrolysis out of the cytoplasm. Certain bacterial and archaeal mPPases can in parallel transport Na+ via a hypothetical "billiard-type" mechanism, also involving the hydrolysis-generated proton. Here, we present the functional evidence supporting this coupling mechanism. Rapid-quench and pulse-chase measurements with [P-32]pyrophosphate indicated that the chemical step (pyrophosphate hydrolysis) is rate-limiting in mPPase catalysis and is preceded by a fast isomerization of the enzyme-substrate complex. Na+, whose binding is a prerequisite for the hydrolysis step, is not required for substrate binding. Replacement of H2O with D2O decreased the rates of pyrophosphate hydrolysis by both Na+- and H+-transporting bacterial mPPases, the effect being more significant than with a non-transporting soluble pyrophosphatase. We also show that the Na+-pumping mPPase of Thermotoga maritima resembles other dimeric mPPases in demonstrating negative kinetic cooperativity and the requirement for general acid catalysis. The findings point to a crucial role for the hydrolysis-generated proton both in H+-pumping and Na+-pumping by mPPases

ПЕРСПЕКТИВНОСТЬ ИНТРОДУКЦИИ НЕТРАДИЦИОННЫХ ПЛОДОВЫХ, ЯГОДНЫХ И ОВОЩНЫХ КУЛЬТУР В УСЛОВИЯХ ДАГЕСТАНА (XI МЕЖДУНАРОДНАЯ НАУЧНО-МЕТОДИЧЕСКАЯ КОНФЕРЕНЦИЯ «ИНТРОДУКЦИЯ, СОХРАНЕНИЕ И ИСПОЛЬЗОВАНИЕ БИОЛОГИЧЕСКОГО РАЗНООБРАЗИЯ КУЛЬТУРНЫХ РАСТЕНИЙ» 10-14 ИЮНЯ 2014

June 9-13, 2014 in Makhachkala hosted XI International scientific-methodical conference on the theme: «Introduction, conservation and use of biological diversity of cultivated plants», organized by FGBNU VNIISSOK, Dagestan Research Institute for Agriculture and GBS DSC RAS. The conference was attended by scientists from Russia, CIS and foreign countries. Due to the conference Dagestan turned out to be a prime location for the cultivation of both traditional and non-traditional plants with a high content of biologically active substances, as well as a training ground for resistance tests because of the combination of mountain and plain zones.9-13 июня 2014 года в Махачкале прошла XI Международная научно-методическая конференция на тему: «Интродукция, сохранение и использование биологического разнообразия культурных растений», организованная ФГБНУ ВНИИССОК, Дагестанским НИИСХ и ГБС ДНЦ РАН. В конференции приняли участие ученые России, ближнего и дальнего зарубежья. По итогам конференции принято заключение, что Дагестан является благоприятным регионом для возделывания как традиционных, так и нетрадиционных растений с повышенным содержанием БАВ, а так же полигоном для испытаний на устойчивость, благодаря сочетанию двух зон: горной и равнинной

The effect of differences in the third domain of the glycoprotein E of tick-borne encephalitis virus of the Far Eastern, Siberian and European subtypes on the binding of recombinant D3 proteins with a chimeric antibody

Currently, a therapeutic drug based on recombinant antibodies for the prevention and treatment of tick-borne encephalitis virus (TBEV) is developed in ICBFM SB RAS, and the chimeric antibody ch14D5 is considered as one of the key components of this drug. It was previously shown that this antibody is directed to the domain D3 of the glycoprotein E of TBEV. It was previously shown that this antibody is able to protect mice from the European subtype of TBEV, strain “Absettarov”, and the presence of virus-neutralizing activity against the Far Eastern subtype of TBEV, strain 205 was also shown for this antibody. However, it remains unclear whether this antibody exhibits selectivity for different subtypes of TBEV. The aim of this study was to investigate the effect of amino acid sequence differences of recombinant D3 domains derived from the glycoprotein E of TBEV of the Far Eastern, Siberian and European subtypes on the binding of the protective antibody ch14D5 to these proteins. Using Western blot analysis and surface plasmon resonance, it was shown that ch14D5 antibody has the highest affinity (KD= 1.7±0.5 nM) for the D3 domain of the TBEV of the “Sofjin-Ru” strain belonging to the Far Eastern subtype of the virus. At the same time, the affinity of ch14D5 antibody for similar D3 proteins derived from “Zausaev”, “1528-99” and “Absettarov” strains of the Siberian and European subtypes of TBEV was noticeably lower (KD= 25±4, 300±50, 250±50 nM, respectively). In addition, information about the spatial arrangement of amino acid residues that are different for the studied recombinant proteins indicates that the epitope recognized by the ch14D5 antibody is in close proximity to the lateral ridge of D3 domain of E glycoprotein

Development of a stable eukaryotic strain producing fully human monoclonal antibody on the basis of the human antibody against ectromelia virus

Fully­human antibodies have a great therapeutic importance; however, the development of stable strains providing a high level of production of full­size antibodies is a challenging task, as antibody molecules contain two types of polypeptide chains. To develop the producing strain, random integration of the plasmid containing the gene encoding the target protein into the genome of the host cells is commonly used. The aim of this study was the development of an original expression system, using gene targeting to integrate the gene encoding the fully­human antibody into the transcriptionally active region of the genome of eukaryotic suspension cells CHO­S. To develop a stable strain, the cassette vector plasmid pCDNA5/FRTDHFR­CH­CL containing the site of homologous recombination and the genes encoding heavy and light chains of the fully human antibody of the IgG1/kappa class was constructed at the first step. Notably, DNA of the plasmid pCDNA5/FRT­DHFR­CH­CL was organized in such a way that the restriction sites for rapid cloning of DNA fragments encoding the variable domains of heavy and light chains were inserted upstream of the sequences encoding constant domains of the heavy and light chains of the antibody. Secondly, DNA fragments encoding the variable domains of the heavy and light chains of antibody against orthopoxvirus protein p35 were inserted into the pCDNA5/FRT­DHFRCH­CL cassette plasmid. Then, CHO­S/FRT cells, which contain the FRT­site for homologous recombination and are able to produce green fluorescence protein GFP, were transfected with the constructed plasmid. After the insertion of the target genes into the FRT­site, GFP production was supposed to stop. Using this selection system, a stable clone producing target antibody fh8E was selected with the level of production of about 100 μg/ml. The binding affinity of purified antibody fh8E with the targeted protein, measured by surface plasmon resonance, was 12 nM. In addition, antibody fh8E demonstrated anti­vaccinia virus activity in the plaque reduction neutralization test in vitro

Beta herpes viruses in patients with cutaneous graft-versus-host disease

Graft-versus-host disease is a major complication of allogenic bone marrow transplantation. Beta herpes viruses’ role in pathogenesis and prognosis of graft-versus-host disease is significant. Objective: To study beta herpes viruses evidence in skin biopsy specimens in graft-versus-host disease patients. Material and methods: Clinical features of 50 patients with cutaneous graft-versus-host disease were examined. Cytomegalovirus and human herpes virus 6 DNA detection in skin biopsy specimens was performed by PCR. Results: Correlation between human herpes virus 6 and poor prognosis in graft-versus-host disease patients has been found. Conclusion: Beta herpes virus detection after allogenic bone marrow transplantation could be important in prognosis of graft-versus-host disease

МОЛЕКУЛЯРНЫЕ МЕХАНИЗМЫ, ОПОСРЕДУЮЩИЕ РАЗВИТИЕ ХОЛАНГИОКАРЦИНОМЫ В ХОДЕ ХРОНИЧЕСКОЙ ИНВАЗИИ ПЕЧЕНОЧНЫМИ СОСАЛЬЩИКАМИ

Cholangiocarcinoma is a malignant tumor, characterized by poor prognosis and a low five-year survival rate. There is a clear correlation between the incidence of opisthorchiasis and high incidence of cholangiocarcinoma in South-East Asia. Liver flukes Clonorchis sinensis and Opisthorchis viverrini are I class carcinogens. There are some endemic regions of opisthorchiasis In the Russian Federation. The most important factor that leads to carcinogenesis during liver fluke infection is chronic inflammation. This review article focuses on the communication of chronic inflammation caused by invasion of liver flukes and cholangiocarcinoma. This paper summarizes the current knowledge about the risk factors for cholangiocarcinoma, as well as knowledge about the molecular aspects of the induction of carcinogenesis by liver flukes.Холангиокарцинома характеризуется неблагоприятным прогнозом и низкой пятилетней выживаемостью. Прослеживается связь между заболеваемостью описторхозом и высокой частотой возникновения холангиокарциномы в странах Юго-Восточной Азии. Печеночные сосальщики Clonorchis Sinensis и Opisthorchis viverrini являются канцерогенами I класса. Известно, что в Российской Федерации также существует несколько эндемических очагов заболеваемости описторхозом. Важнейшим фактором, который обусловливает канцерогенез при описторхозной инвазии, является хроническое воспаление. В обзорной статье основное внимание уделено связи хронического воспаления, вызванного инвазией печеночных сосальщиков, и холангиокарциномы. Обобщены имеющиеся знания о факторах риска развития холангиокарциномы, а также данные о молекулярных аспектах индукции канцерогенеза печеночными сосальщиками

СОДЕРЖАНИЕ ПОЛИФЕНОЛОВ, АСКОРБИНОВОЙ КИСЛОТЫ, КАРОТИНОИДОВ И АНТИОКСИДАНТНЫЕ СВОЙСТВА ПЛОДОВ CAPSICUM CHINENSE JACQ.

Capsicum chinense Jacq. is the rich source of phytochemical substances, such as vitamin C, phenolic compounds, carotenoids, and occupies a significant place in the cooking of many peoples. The pigment composition, the amount of carotenoids, ascorbic acid and their contribution to the formation of antioxidant properties were studied. The total content of carotenoids and red/yellow pigments varied greatly among the accessions of C. chinense. The highest content of carotenoids 0.581 mg/g was observed in the varieties ‘Ognennaya Deva’ with the red fruit color in the phase of biological ripeness, where red pigments accumulated 56% more than in yellow ones. Fruits of accessions ‘Kolokolchik’ only accumulated yellow pigments, 0.318 mg/g. Presence of significant amount, 2.03 times as much red pigments as yellow ones in the variety ‘Trinidad Scorpion Chocolate’, with protein complex could cause the formation of a brown fruit color. Another tendency was observed in ‘Trinidad Dglahou’. The yellow pigment is 1.5 times as much amount as red, as estimated 0.119 mg/g of yellow and 0.077 mg/g of red pigment. The ascorbic acid content did not depend on the fruit color. Fruits of the ‘Ognennaya Deva’ accumulated the highest amount of ascorbic acid, 301 mg%. The maximum total antioxidant content, was detected in the variety ‘Ognennaya Deva’ 2.65 (TAC, mg.eq. GA/g). Analysis of the dependence of the content of thermo stable antioxidants to the total ones showed the contribution of the unstable antioxidants to thermal effects (particularly ascorbic acid) on the level of 16%, on average. A comparative assessment of the pungency level and quantitative capsaicin content in the fruit was carried out by organoleptic, spectrophotometric and HPLC methods. The intervals of observed concentrations of capsaicin were 1.0-7.5 mg/g of dry weight (HPLC), the level of pungency on the Scoville scale was 17440-153120 SHU. There was a direct correlation between the organoleptic assessment of the pepper fruit pungency and capsaicin content studied by spectrophotometry and HPLC methods.Плоды Capsicum chinense Jacq. являются богатым источником фитохимических веществ, такие как витамин С, фенольные соединения, флавоноиды и каротиноиды, и занимают значительное место в кулинарии многих народов. Изучен пигментный состав, определена сумма каротиноидов, аскорбиновой кислоты и их вклад в формирование антиоксидантных свойств. Среди изученных образцов C. chinense как общее содержание каротиноидов, так и красных, и желтых пигментов варьирует значительно. Самое высокое содержание каротиноидов наблюдается у сорта Огненная дева с красной окраской плодов в фазе биологической спелости – 0,581 мг/г, у которого красных пигментов накапливается на 56% больше, чем желтых. В плодах образца Колокольчик накапливаются только желтые пигменты (0,319мг/г). Наличие значительного количества красных пигментов, в 2,03 раза превышающих количество желтых у сорта Trinidad Scorpion Chocolate, в комплексе с белками, могли стать причиной формирования коричневой окраски. У образца Trinidad Dglahou наблюдается другая тенденция. Количество желтых пигментов в 1,5 раза больше, чем красных: 0,118 мг/г желтых и 0,077 мг/г красных пигментов. Содержание аскорбиновой кислоты не зависит от окраски плода. В плодах сорта Огненная дева накапливается наибольшее количество аскорбиновой кислоты – 301мг%. Максимальное суммарное содержание антиоксидантов (ССА, мг.экв. ГК/г) выявлено у сорта Огненная дева – 2,65. Анализ зависимости содержания термостабильных антиоксидантов от общего их количества показал вклад неустойчивых к термическому воздействию антиоксидантов (в первую очередь аскорбиновой кислоты) на уровне, в среднем, 16%. Проведена сравнительная оценка уровня остроты и количественное содержание капсаицина в плодах органолептическим, спектрофотометрическим и ВЭЖХ методами. Интервал наблюдаемых концентраций капсаицина составил 1,0-7,5 мг/г сухой массы (ВЭЖХ), уровень остроты по шкале Сковилля – 17440-153120 SHU. Установлена прямая корреляция между органолептической оценкой остроты плодов перца и содержанием капсаицина, определенным спектрофотометрическим и ВЭЖХ методами