Development of a novel genus-specific real-time PCR assay for detection and differentiation of Bartonella species and genotypes

The genus Bartonella includes numerous species with varied host associations, including several that infect humans. Development of a molecular diagnostic method capable of detecting the diverse repertoire of Bartonella species while maintaining genus specificity has been a challenge. We developed a novel real-time PCR assay targeting a 301-bp region of the ssrA gene of Bartonella and demonstrated specific amplification in over 30 Bartonella species, subspecies, and strains. Subsequent analysis of ssrA sequences was sufficient to discriminate Bartonella species and provided phylogenetic data consistent with that of gltA, a commonly used gene for differentiating Bartonella genotypes. Using this assay, we identified Bartonella DNA in 29% and 47% of blood specimens from elk in Wyoming and cattle in the Republic of Georgia, respectively. Sequence analysis of a subset of genotypes from elk specimens revealed a cluster most closely related to Bartonella capreoli, and genotypes from cattle were identified as Bartonella bovis, both Bartonella species commonly found in wild and domestic ruminants. Considering the widespread geographic distribution and infectivity potential to a variety of hosts, this assay may be an effective diagnostic method for identification of Bartonella infections in humans and have utility in Bartonella surveillance studies.     

Bartonella tamiae sp. nov., a newly recognized pathogen isolated from three human patients from Thailand

Three strains of a novel Bartonella species (Bartonella tamiae) were isolated from human patients from Thailand. Sequence analysis of six chromosomal regions (16S rRNA, gltA, groEL, ftsZ, rpoB, and the intergenic spacer region) and phenotypical analysis supported the similarity of the three strains and placed them within the genus Bartonella separately from previously described species.     

Serological response to Bartonella species in febrile patients from Nepal

The Bartonella-associated illnesses are spread world-wide and involve a broad spectrum of signs and symptoms in humans. Several Bartonella species have been shown to be responsible for cases of febrile illnesses. Little information exists on distribution of Bartonella species and their role in human diseases in Nepal. Our preliminary study, a retrospective serological survey of archived specimens, suggests that Bartonella antibodies are prevalent among febrile patients in the Kathmandu Valley of Nepal.     

Intense pulsed light vs. pulsed‐dye laser in the treatment of facial acne: a randomized split‐face trial

Background Various laser and light therapy have been increasingly used for the treatment of acne vulgaris. Patients and methods Twenty patients with facial acne were treated using intense pulsed light (IPL) on one side of the face and pulsed dye laser (PDL) on the other to compare the efficacy and safety of IPL and PDL. Treatment was performed 4 times at 2‐week intervals. Treatment effectiveness was determined using lesion counts, acne severity, patient subjective self‐assessments of improvement, and histopathological examinations, which included immunohistochemical staining for transforming growth factor‐β (TGF‐β). Results Numbers of total acne lesions decreased following both treatments. For inflammatory lesions such as papules, pustules and nodules, IPL‐treated sides showed an earlier and more profound improvement than PDL‐treated sides. However, at 8 weeks after the 4th treatment, a rebound aggravation of acne was observed on IPL‐treated sides. On the contrary, PDL produced gradual improvements during the treatment sessions and these improvements lasted 8 weeks after the 4th treatment. Non‐inflammatory lesions as open and closed comedones also showed improvement following both treatments and PDL‐treated sides showed better improvement as the study proceeded. Histopathological examinations showed amelioration in inflammatory reactions and an increase in TGF‐β expression after both treatments, which were more prominent for PDL‐treated sides. Conclusion Both PDL and IPL were found to treat acne effectively, but PDL showed a more sustained effect. TGF‐β might play a key role in the resolution of inflammatory acne lesions.     

Human Lymphadenopathy Caused by Ratborne Bartonella,Tbilisi, Georgia

Lymphadenopathy and fever that developed in a woman in Tbilisi, Georgia, most likely were caused by a ratborne Bartonella strain related B. tribocorum and B. elizabethae. The finding suggests that this Bartonella strain could be spread by infected rats and represents a potential human risk.     

HPLC法同时测定注射用甲氧异腈中FSA和MIBI的含量

目的 :采用RP HPLC法同时测定了注射用甲氧异腈中两种主要成份二氧硫脲 (FSA)和四 (甲氧基异丁基异腈 )络铜 (I)氟硼酸盐 (MIBI)的含量。方法 :以甲醇、磷酸二氢铵 ( 80∶2 0 )为流动相 ,检测波长 2 1 5nm ,HPLC法测定含量。结果 :试验表明 ,FSA和MIBI在 2 5～ 1 2 5μg/ml,50～ 2 50 μg/ml范围内呈良好的线性关系 ,回归方程分别为Y =0 .2 3 0 .0 2X(r =0 .994 6) ,Y =-2 .1 9 0 .0 3X(r =0 .9998) ,相对标准偏差分别为 1 .2 5%和 0 .4 2 %。结论 :该方法简便、准确、可靠     

Synthesis and biological activities of fluorescent acridine-containing HIV-1 nucleocapsid proteins for investigation of nucleic acid-NCp7 interactions

Specific interactions between the 72-amino acid nucleocapsid protein NCp7 of the human immunodeficiency virus, type 1 and the genomic RNA are essential for virus replication. Studies on the mechanism of action of NCp7 require a direct visualization of its complexes with nucleic acids and the determination of binding affinities. To facilitate these investigations, fluorescent NCp7 derivatives were developed by introduction in the NCp7 sequence of a non-natural amino acid, (S)-β-(9-acridinyl)alanine (Aca) obtained by a chiral synthetic method. Three fluorescent NCp7 derivatives were obtained by introducing this amino acid at different positions. As shown by NMR, the three-dimensional structure of NCp7 is not altered by introduction of Aca. The fluorescent peptides were found to be as potent as their precursors in interacting with nucleic acids and in promoting HIV-1 genomic RNA dimerization. Moreover, because of their fluorescent properties, these NCp7s can be used at submicromolar concentrations to directly visualize and quantify protein-nucleic acid interactions in solution or after gel electrophoresis. This could facilitate the development of new antiviral agents aimed at inhibiting the functions of NCp7 and studies on the intracellular traffic of NCp7 within the preintegration complex.     

转化生长因子β1在白细胞介素1β诱导大鼠肾小管上皮细胞转分化及大黄素干预中的作用

目的:大黄素对白细胞介素1β诱导NRK52E细胞转分化有显著抑制作用。实验拟进一步观察转化生长因子β1在白细胞介素1β诱导大鼠肾小管上皮细胞-肌成纤维细胞转分化及大黄素抑制作用中的意义。方法:实验于2006-10/2007-05在泸州医学院附属医院免疫实验室完成。⑴实验材料及分组:以培养的大鼠肾小管上皮细胞株(NRK52E)为观察对象,按如下分组分别添加不同处理因素:①对照组:仅加入体积分数为0.05小牛血清的高糖DMEM培养基。②白细胞介素1β诱导组:加含白细胞介素1β终浓度为10μg/L的高糖DMEM培养基。③SB431542阻断组:加含白细胞介素1β终浓度为10μg/L及SB431542终浓度为10μmol/L的高糖DMEM培养基。④白细胞介素1β 大黄素组:同时加分别含白细胞介素1β终浓度为10μg/L及大黄素终浓度为25mg/L的高糖DMEM培养基。⑵实验评估:培养48h后用倒置相差显微镜观察细胞形态,细胞免疫化学染色法检测肌酸激酶、α-平滑肌肌动蛋白及转化生长因子β1的表达。结果:①白细胞介素1β可诱导部分细胞由卵圆形转变为梭形,且肌酸激酶表达减弱(P<0.01),α-平滑肌肌动蛋白及转化生长因子β1表达显著增强(P<0.01)。②SB431542特异性抑制转化生长因子β1作用后,白细胞介素1β诱导的细胞形态改变受抑,同时肌酸激酶表达增强(P<0.01),α-平滑肌肌动蛋白表达减弱(P<0.01),但转化生长因子β1的表达却无明显变化。③大黄素对白细胞介素1β诱导的细胞形态改变及肌酸激酶、α-平滑肌肌动蛋白的表达有明显抑制作用,其抑制作用与SB431542的作用相比无显著差异;同时,大黄素对白细胞介素1β诱导的转化生长因子β1的表达也有明显抑制作用(P<0.01)。结论:转化生长因子β1可能介导了白细胞介素1β诱导大鼠肾小管上皮细胞-肌成纤维细胞转分化,并参与了大黄素抑制白细胞介素1β诱导大鼠肾小管上皮细胞转分化的作用。