Genetic engineering of peptide hormones : II. Possible polymorphism of preprolactin in cattle. Data of molecular cloning

Primary structure is determined of an insertion of a clone isolated from the library of hypophyseal cDNA of cattle by hybridization with a probe specific for prolactin. Analysis of nucleotide sequences showed that in the process of cloning, reorganization occurred in structure of preprolactin cDNA, including an inversion of the 5'-terminal and deletion of the central section of cDNA. Nevertheless, from structure of cDNA, nucleotide sequences can be deduced of extended 5'- and 3'-terminal sections of preprolactin mRNA in cattle with lengths of 257 and 551 nucleotide residues, respectively. When these sequences are compared to those established previously, some differences were found in primary structure. The most important of them is the presence of an additional codon which codes alanine at the position (-22) of the signal peptide. It is suggested that heterogeneity of preprolactin mRNA of cattle in the section coding the signal peptide is the result of alternative splicing, as was shown for preprolactin mRNA in rats

Identification of Bacterial Clones Encoding Bovine Caseins by Direct Immunological Screening of the cDNA Library

A sensitive immunoassay was used to identify recombinant plasmids carrying cDNA fragments of bovine caseins in the cDNA library from bovine mammary gland mRNA. Colonies grown on nitrocellulose filters were lysed in situ and proteins from the lysates were blotted onto CNBr-activated cellulose filter paper. Antigens covalently bound to CNBr-activated paper or bound to nitrocellulose filters were detected by reaction with antiserum to caseins, followed by ^125I-labelled Staphylococcus aureus protein A and autoradiography. Six clones were found positive among 5400 of the cDNA library: 3-A1, 3-B2, 3-B5, 3-H7, 2-A5 and 2-C9. The molecular weights of chimeric pre-β-lactamase: casein proteins synthesized in Escherichia coli were estimated by immunoblotting. Colony hybridization and nucleotide sequence analysis showed that clone 3-B5 contained a cDNA fragment of bovine χ-casein, clone 3-H7 contained a cDNA fragment of β-casein, while clones 2-A5 and 2-C9 carried cDNA fragments of α_si-casein

Identification of Bacterial Clones Encoding Bovine Caseins by Direct Immunological Screening of the cDNA Library

A sensitive immunoassay was used to identify recombinant plasmids carrying cDNA fragments of bovine caseins in the cDNA library from bovine mammary gland mRNA. Colonies grown on nitrocellulose filters were lysed in situ and proteins from the lysates were blotted onto CNBr-activated cellulose filter paper. Antigens covalently bound to CNBr-activated paper or bound to nitrocellulose filters were detected by reaction with antiserum to caseins, followed by ^125I-labelled Staphylococcus aureus protein A and autoradiography. Six clones were found positive among 5400 of the cDNA library: 3-A1, 3-B2, 3-B5, 3-H7, 2-A5 and 2-C9. The molecular weights of chimeric pre-β-lactamase: casein proteins synthesized in Escherichia coli were estimated by immunoblotting. Colony hybridization and nucleotide sequence analysis showed that clone 3-B5 contained a cDNA fragment of bovine χ-casein, clone 3-H7 contained a cDNA fragment of β-casein, while clones 2-A5 and 2-C9 carried cDNA fragments of α_si-casein

Enhanced four-wave mixing via elimination of inhomogeneous broadening by coherent driving of quantum transition with control fields

We show that atoms from wide velocity interval can be concurrently involved in Doppler-free two-photon resonant far from frequency degenerate four-wave mixing with the aid of auxiliary electromagnetic field. This gives rise to substantial enhancement of the output radiation generated in optically thick medium. Numerical illustrations addressed to typical experimental conditions are given.Comment: LaTeX2e, hyperref, 7 pages, 5 figures, to appear in PRA 1 august 200

CVD graphene sheets electrochemically decorated with “core-shell” Co/CoO nanoparticles

The paper reports on the first successful fabrication of Co-graphene composites by electrochemical deposition of Co nanoparticles (NPs) on the sheets of twisted graphene. Characterization of the surface morphology and element mapping of twisted graphene decorated with Co NPs by transmission and scanning electron microscopy in combination with the energy-dispersive X-ray spectroscopy reveals the formation of isolated quasi-spherical oxidized Co NPs with the mean diameter h di _ 220 nm and core-shell structure. X-ray photoelectron spectroscopy indicates that the core of deposited NPs consists of metal Co while the shell is CoO. Composite Co-graphene samples containing core-shell NPs reveal an exchange bias field up to 160 Oe at 4 K as detected by vibrating sample magnetometry after the field cooling procedure

ДИНАМИКА СОСТОЯНИЯ УГЛЕВОДНОГО ОБМЕНА ПОСЛЕ РЕНАЛЬНОЙ ДЕНЕРВАЦИИ У БОЛЬНЫХ РЕЗИСТЕНТНОЙ АРТЕРИАЛЬНОЙ ГИПЕРТОНИЕЙ В СОЧЕТАНИИ С САХАРНЫМ ДИАБЕТОМ 2-ГО ТИПА

The aim of the study was to evaluatetheglycemic control dynamics depending on degree of blood pressure (BP) reduction and dynamic of TNF-α after 6 and 12 months of Tran catheter renal denervation (TRD) of patients with true resistant hypertension (RH) and type 2 diabetes mellitus (T2DM). Material and methods. Thirty two essentially hypertensive patients with type 2 diabetes mellitus (T2DM) and resistant hypertension were included in single-arm prospective interventional study. Office BP measurement, ambulatory 24-h BP, renal Doppler ultrasound and assessment of renal function (proteinuria, creatinine, eGFR), HbА1c and fasting plasma glucose (FPG) levels, activity of TNF-α were performed at baseline and 6 and 12 months after TRD. On average, patients were taking 4 (3–6) antihypertensive drugs. None of the patients changed the antihypertensive treatments during follow-up. A 6 months follow-up was completed by 27 patients (43–75 years old, 14 male), 12 months follow-up was completed by 26 patients. Results. Renal denervation significantly reduced the systolic office BP (SBP) as well as 24-h SBP (– 27.2/–10.7 mm Hg and–13.4/–10 mm Hg, respectively, p < 0.01 after 6-month follow-up, and –31,7/–12,8 mmHg and –13.4/–10 mm Hg, respectively, p < 0.01 after 12-month follow-up) without any negative effect on renal function. The number of responders with reduction of SBP >10 mmHg according to ABPM were 56% (15/27) after 6-month and 61.5% (16/26) after 12-month follow-up. There were significant reduction of the average HbA1c levels (from (6.9 ± 1.8)% to (5.8 ± 1.5)%, p = 0.04) and nonsignificant decreasing of FPG levels (from 8.7 ± 2.8 to 7.7 ± 2.1 mmol/L, p = 0.07) after 6-month followup. Conspicuously, the responders according to ABPM had significantly higher mean dynamics of HbA1c than the non-responders after 6-month follow-up (–2.4 ± 1.9 and –0.1 ± 0.8%, p = 0.02, respectively) as well as after 12-month follow-up (–0.12 ± 0.98 and 1,26 ± 1.11%, p = 0.04 for HbA1c, and – 0.89 ± 1.9 и 0.85 mmol/L ± 1.19, p = 0.02 for FPG levels). There were significant decreasing of TNF-α after 12-monthfollow-up (from 2.21 (1.54–3.65) to 1.4 (1.11–1.47pg/ml), p = 0.007), without relation to BP and HbA1c dynamics, and response to TRD. There were not the correlations between dynamics of HbA1c and FPG levels with BP reduction and change of TNF-α after 12-month follow-up. Conclusions. Renal denervation of patients with true resistant hypertension and diabetes mellitus type 2 after 6 and 12 months was followed by improved glycemic control, BP reduction and decreasing of mean levels of TNF-α. Glycemic control improvement after the renal denervation was more expressive in the responders. Цель исследования – оценить изменение состояния углеводного обмена через 6 мес и 1 год после ренальной денервации (РД) в зависимости от степени антигипертензивного эффекта и динамики фактора некроза опухолей α (ФНО-α) у больных резистентной артериальной гипертонией (РАГ) в сочетании с сахарным диабетом (СД) 2-го типа (СД-2).Материал и методы. Ренальная денервация почечных артерий проведена у 32 больных истинно резистентной АГ в сочетании с СД-2. Полугодовой период наблюдения закончили 27 пациентов (14 мужчин и 13 женщин (возраст 43–75 лет)), годовое наблюдение – 26 человек. Всем больным проводили общеклиническое обследование, измерение офисного артериального давления (АД), 24-часовое амбулаторное мониторирование АД (АМАД), допплерографию почечных артерий, оценивали состояние углеводного обмена (базальная гликемия, гликированный гемоглобин (HbA1c), концентрацию в плазме крови ФНО-α и почечную функцию (протеинурия, креатинин сыворотки крови, расчетная скорость клубочковой фильтрации (по формуле MDRD). Пациенты получали в среднем 4 (3–6) антигипертензивных препарата. Антигипертензивная и сахароснижающая терапия оставались стабильными в течение всего периода наблюдения. Результаты. Через 6 мес после РД отмечался значимый антигипертензивный эффект (–27,2/–10,7 мм рт. ст. для офисного АД и –13,4/–10,0 мм рт. ст. для АД–24-ч, p < 0,01), остававшийся стабильным в течение всего года (–31,7/–12,8 мм рт. ст. и –13,4/–10,0 мм рт. ст., соответственно,p < 0,01). Снижение систолического АД (САД) более 10 мм рт. ст от исходных значений (группа респондеров) по результатам АМАД через 6 мес зарегистрировано у 56% больных (15 из 27 пациентов), через год – 61,5% (16 из 26 человек). Через 6 мес после вмешательства установлено статистически значимое уменьшение среднего уровня HbA1c (от (6,9 ± 1,8) до (5,8 ± 1,5)%, p = 0,04) и тенденция к снижению базальной гликемии (от 8,7 ± 2,8 до (7,7 ± 2,1) ммоль/л, p = 0,07). При этом снижение HbA1c было более выраженным у респондеров (по САД–24 ч), чем у нереспондеров  (–2,4 ± 1,9 и –0,1 ± 0,8 соответственно, p = 0,02). Различие динамики HbA1c между респондерами и нереспондерами cохранялось и через год наблюдения (–0,1 ± 1,0 и 1,3 ± 1,1, p = 0,04), также как и динамики базальной гликемии (–0,9 ± 1,9 и 0,8 ± 1,2, p = 0,02). Через год после РД было обнаружено статистически значимое снижение уровня ФНО-α (от 2,21 (1,54–3,65) до 1,40 (1,11–1,47) пг/мл),p = 0,007), не зависящее от выраженности снижения АД и ответа на вмешательства. Прямой связи между снижением показателей углеводного обмена и динамикой концентрации ФНО-α, а также степенью антигипертензивного эффекта через год также не отмечено. Выводы. Ренальная денервация у больных резистентной АГ, ассоциированной с СД-2, оказывала благоприятное влияние на состояние углеводного обмена на фоне выраженного и стабильного в течение года антигипертензивного эффекта, а также снижения активности ФНО-α. При этом статистически значимое улучшение гликемического контроля отмечалось при снижении среднесуточного систолического АД более 10 мм рт. ст. от исходных значений

On the origin of the H1N1 (A/USSR/90/77) influenza virus

The influenza virus H1N1 (the A/USSR/90/77 strain) that reappeared in 1977 after the H1N1 influenza viruses had disappeared from the human population, is compared with the A/FM/1/47 and the A/FW/1/50 influenza viruses by the method of oligonucleotide mapping of individual segments of the viral RNAs. Seven genes of the A/USSR/90/77 virus appear to be very similar to the corresponding genes of the A/FW/1/50 virus, whereas the gene coding for the M protein displays considerable homology to the corresponding gene of the A/FM/1/47 virus. The data demonstrate that the A/USSR/90/77 strain is a recombinant virus