Anti-measles DNA-immunization in experiment: immunogenicity and safety

The plasmid DNA pCDNA3.1-H encoding the N-terminal sequence of the measles hemagglutinin (H) protein was constructed. Virus-specific particles (VSP) containing the plasmid DNA pCDNA3.1-H coated by the spermidine-polyglucin complex. The mice were immunized by VSP. ELISA, HAIT and immunoblot showed a shaping specific humoral response. The sera of immunized animals were proven to neutralize the wild strain of the NOV96 measles virus. The formation of the specific cell immunity was confirmed by erythrocyte proliferation assay and ELISpot. PCR was used to detect the presence of the plasmid DNA in different intestines and tissues of animals after a single immunization. It was not detected at any time interval in the brain, liver, thymus and blood. And it was present on days 7 and 14 in the red bone marrow, spleen, muscular tissue, lungs and fatty tissue. On day 21 the plasmid DNA was not detected in any of the investigated organs.     

AE9C90CB: a novel, bladder-selective muscarinic receptor antagonist for the treatment of overactive bladder

Background and purpose:

AE9C90CB (N- [(1R, 5S, 6R)-3-azabicyclo [3.1.0] hex-6-ylmethyl]-2-hydroxy-N-methyl-2, 2-diphenylacetamide), a novel muscarinic receptor antagonist, was synthesized for the treatment of overactive bladder. Here we describe the in vitro and in vivo profiles of AE9C90CB for action in bladder over salivary gland and compare it with four agents already in clinical use (tolterodine, oxybutynin, solifenacin and darifenacin).

Experimental approach:

Radioligand binding assay and isolated tissue-based functional assay were used to evaluate affinity, potency, and receptor subtype selectivity of compounds. Inhibition of carbachol-induced increase in intravesicular pressure and salivary secretion were measured in anaesthetized rabbits to assess the functional selectivity.

Key results:

In vitro radioligand binding study using human recombinant muscarinic receptors showed that AE9C90CB had greater affinity for M₃ muscarinic receptors with pKi of 9.90 ± 0.11 and was 20-fold more selective for M₃ than for M₂ muscarinic receptors. AE9C90CB exhibited an unsurmountable antagonism on rat bladder strips (pK_B, 9.13 ± 0.12). In anaesthetized rabbits after intravenous administration, AE9C90CB dose dependently inhibited carbachol-induced increase in intravesicular pressure and salivary secretion, and exhibited functional selectivity for urinary bladder over salivary gland which was ninefold better than that of oxybutynin.

Conclusions and implications:

We have identified AE9C90CB, a compound exhibiting moderate selectivity for M₃ over M₂ receptors but greater selectivity for urinary bladder over salivary gland than oxybutynin, tolterodine, solifenacin and darifenacin. Therefore, AE9C90CB may be a promising compound for the treatment of overactive bladder with reduced potential to cause dry mouth than currently available antimuscarinic drugs.     

A synergistic effect of a combined bivalent DNA-protein anti-HIV-1 vaccine containing multiple T- and B-cell epitopes of HIV-1 proteins

Immunogenic properties of the combined vaccine CombiHIVvac, comprising polyepitope HIV-1 immunogens, one being the artificial polyepitope protein TBI, containing the T- and B-cell epitopes from Env and Gag proteins, and the DNA vaccine construct pcDNA-TCI coding for the artificial protein TCI, carrying over 80 T-cell epitopes (both CD4+ CTL and CD8+ Th) from Env, Gag, Pol, and Nef proteins, are studied in this work. The data reported demonstrate clearly that a combination of two B- and T-cell immunogens (TBI and TCI) in one construct results in a synergistic increase in the antibody response to both TBI protein and the proteins from HIV-1 lysate. The level of antibodies induced by immunization with the constructs containing either immunogen alone (TBI protein or the plasmid pcDNA-TCI) was significantly lower as compared to that induced by the combined vaccine. The analysis performed suggests that the presence of CD4+ T-helper epitopes, which can be presented by MHC class II, in the protein TCI may be the main reason underlying the increased synthesis of antibodies to TBI protein due to a CD4-mediated stimulation of B-cell proliferation and differentiation.     

The relationship between urinary calcium, sodium, and potassium excretion and the role of potassium in treating idiopathic hypercalciuria

OBJECTIVES: 1) To evaluate the relationships between urinary sodium (UNa), potassium (UK), and calcium (UCa) excretion in the pediatric population; and 2) to determine the effect of increasing potassium intake in patients with idiopathic hypercalciuria and investigate whether this intervention can be offered as another mode of therapy in this patient population. DESIGN: Prospectively, we determined UNa, UK, UCa, and creatinine (Cr) concentrations in randomly collected urine samples from children on initial evaluation for urinary frequency, dysuria, hematuria, enuresis, or kidney stones to identify children with hypercalciuria. SETTING: The outpatient renal clinic of an academic hospital. PARTICIPANTS: Twenty-three black children (13 girls and 10 boys) and 77 white children (44 girls and 33 boys) 3.92 to 16.67 years of age. INTERVENTIONS: Eleven children with hypercalciuria were given potassium supplementation or placed on a high-potassium diet for at least 2 weeks. OUTCOME MEASURES: UNa to UK, UNa to Cr, UK to Cr, and UCa to Cr ratios were calculated from measured levels of urinary minerals. These were repeated in 11 hypercalciuric patients after 2 weeks of increased potassium intake. RESULTS: A total of 100 urine samples were analyzed. The UCa/Cr ratio in blacks 0.04 +/- 0.06 (mean +/- standard deviation) was significantly lower than in whites 0.16 +/- 0.12. There were 21 hypercalciuric white children versus only 1 black child. Linear regression analysis revealed a positive direct correlation between UNa/Cr and UCa/Cr in all 100 subjects and in whites alone but not in blacks. An inverse relationship existed between UK/Cr and UCa/Cr in all subjects and in whites and showed a strong trend in blacks. A marked direct relationship was found between UNa/K and UCa/Cr in all subjects (r = .43) as well as in whites (r = .59) and blacks (r = .49). One black child and 10 white hypercalciuric children were treated with "extra" K for at least 2 weeks. The UNa/K decreased from 4.73 +/- 2.28 to 1.98 +/- 1.09, and the UCa/Cr decreased from 0. 31 +/- 0.10 to 0.14 +/- 0.07, with resolution or improvement of the patients' symptoms. CONCLUSIONS: In our patient population with urinary symptoms, the UCa/Cr ratio in black children is lower and hypercalciuria less common than in white children. In both white and black populations, the UNa/K ratio had the strongest association with the UCa/Cr ratio, indicating an opposing role of UNa and UK on the UCa/Cr ratio. Increased potassium intake was found to be beneficial for hypercalciuric children by decreasing the UNa/K ratio and, consequently, the UCa/Cr ratio.     

Construction and study of antigenic characteristics of recombinant salmonella strain producing TBI protein

A recombinant strain producing TBI protein (artificial protein containing HIV-1 B and T cell epitopes) was constructed on the base of attenuated Salmonella typhimurium SL7207 strain and BALB/c mice were immunized with this strain in a dose of 10(9) live cells orally or rectally. A single immunization with the recombinant salmonella strain induced humoral and cellular immune response to HIV-1; hence, this strain is a promising candidate for vaccine against HIV.     

L一粘附素单抗在缺血再灌注损伤中的作用

目的：中性粒细胞粘附在缺血再灌注损伤中有非常重要的作用。本文用ＳＤ大鼠趾长屈肌缺血再灌注损伤模型,观察Ｌ一粘附素单抗ＬＡＭ１—１１６在缺血再灌注损伤中的作用。方法：３０只ＳＤ大鼠被均分为２组：ＬＡＭ１—１１６组和生理盐水对照组。每只大鼠的一侧趾长屈肌作为正常对照,另外一侧进行 ３ ｈ缺血 ４ ｈ再灌注。结果：ＬＡＭ１— １１６组实验侧的髓过氧化物酶为正常的２倍（２．３±２．２）,生理盐水对照组则为正常的２８倍（２７．５±１１．７）（Ｐ＜０．００１）;ＬＡＭ１—１１６组的湿重比（１．１０± ０．１０）、疲劳肌力（７７． １％±１２．１％）与对照组相比（分别为 １． ２３± ０． １０和 ４９． ７％± ９ ．３％）明显改善（Ｐ＜ ０．０５）;组织学上,ＬＡＭ１—１１６组的中性粒细胞局部浸润显著减少,水肿减轻。结论：通过 Ｌ－粘附素单克隆抗体 ＬＡＭ１— １１６阻断 Ｌ－粘附素的功能,可以有效地降低中性粒细胞在再灌注肌肉中的浸润,防止组织水肿,从而改善肌肉的功能。