Differential effects of caffeine on dihydroxyphenylacetic acid concentrations in various rat brain dopaminergic structures

The behavioural and neurochemical effects of caffeine were examined in rats. The intraperitoneal administration of different doses of caffeine significantly decreased DOPAC concentrations in striatum, hypothalamus and frontal cortex, but increased them in nucleus accumbens. These observations suggest that the effects of caffeine on the central nervous system (cns) are at least partially mediated through an interaction with the dopaminergic system.     

Lifelong treatment with gammaglobulin for primary antibody deficiencies: the patients' experiences of subcutaneous self-infusions and home therapy

Primary antibody deficiencies are chronic conditions and the patients usually need lifelong replacement therapy with gammaglobulin to prevent or reduce infections It has been shown that the gammaglobulm can be given safely as subcutaneous infusions, instead of intramuscular injections or intravenous infusions The major aim of this multi-centre study was to investigate the perceptions of the subcutaneous method among patients using it, both in hospital settings and as self-infusions at home The study included 152 patients 89 women, 63 men, mean age 44 years (range 18-76) Data were collected by using questionnaires The patients were found to have a strongly positive attitude towards receiving the replacement therapy as subcutaneous infusions, perceived the method as effective in preventing infections and wished to retain the treatment However, the younger patients found the subcutaneous infusions more uncomfortable and were less determined to continue with the therapy as compared with the older individuals The responsibility for self-infusions at home was accepted by the patients, leading to an increased independence from the health care personnel and to a feeling of flexibility and freedom As these patients have a chronic disease and are in need of lifelong treatment, it is important to discuss the development of structured education and training programmes in which special emphasis is placed on the support of the younger patients It is suggested that Orem's nursing model of self-care may serve as a conceptual framework for nurses working in this specific area of nursing care     

Prevalence and Associations of tcpC,a Gene Encoding a Toll/Interleukin-1 Receptor Domain-Containing Protein,among Escherichia coli Urinary Tract Infection,Skin and Soft Tissue Infection,and Commensal Isolates

TcpC, a new Toll/interleukin-1 receptor domain-containing protein of uropathogenic Escherichia coli involved in the suppression of innate immunity, was found in 2008. The aim of the present study was to determine the prevalence of tcpC and its association with virulence factors and phylogenetic groups among strains from a collection of 212 E. coli isolates from urinary tract and skin and soft tissue infections and 90 commensal E. coli strains.Pathogenic microbes avoid host defenses using a wide array of virulence factors. Escherichia coli strains, even though they are common bacteria of the gut microbiota, can be important pathogens due to the possession of virulence factors (5). Recently, Cirl et al. (1) reported that they found TcpC, a new Toll/interleukin-1 receptor (TIR) domain-containing protein of uropathogenic E. coli that inhibits Toll-like receptor (TLR) and MyD88-specific signaling, thus impairing the innate immune response. They further reported that tcpC homologous sequences were present in about 40% of E. coli isolates from individuals with pyelonephritis, 21% of isolates from individuals with cystitis, 16% of isolates from individuals with asymptomatic bacteriuria, and only 8% of commensal isolates. Their results suggested that TcpC increases the severity of urinary tract infections (UTIs) in humans and provided the first unambiguous evidence that bacterial pathogens interfere with TLR signaling to survive and spread in the human host.The aim of our study was to determine the prevalence of tcpC among 212 extraintestinal E. coli isolates: 100 E. coli isolates from individuals with symptomatic UTIs, 10 E. coli isolates from individuals with asymptomatic UTIs, 102 E. coli isolates from isolates from individuals with skin and soft tissue infections (SSTIs), and 90 E. coli commensal isolates. In addition, we investigated the association of tcpC with the phylogenetic group (groups A, B1, B2, and D; E. coli strains causing extraintestinal infections are known to mainly belong to group B2 and, to a lesser extent, group D, while commensal E. coli strains belong to groups A and B1), as well as with other well-known virulence factors of extraintestinal pathogenic E. coli (ExPEC) strains (cytotoxic necrotizing factor 1 [cnf1], hemolysin [hlyA], P-fimbrial adhesins [papGIII and papGII], S fimbriae [sfaDE], Afa/Dr adhesins [afa/draBC], aerobactin [iucD], and uropathogenic strain-specific protein [usp]). To our knowledge, this is the first investigation of the prevalence of tcpC among E. coli strains causing SSTIs and of the association of tcpC with phylogenetic group as well as virulence factor genes among UTI, SSTI, and commensal E. coli isolates.The extraintestinal E. coli isolates examined in this study were from our previous studies of UTIs (10, 12-14) and SSTIs (9), while the 90 E. coli commensal isolates were isolated for the purposes of this study. The commensal E. coli isolates were isolated as lactose-positive colonies on MacConkey agar plates from the feces of healthy individuals. Indole, methyl red, Voges-Proskauer, and citrate tests were performed to ascertain that the species detected were E. coli. The strains investigated were cultivated in Luria-Bertani medium or agar. Cell lysates of all 302 E. coli isolates were prepared (7) and used in the PCRs. Amplifications were performed in an automated thermal cycler (UNOII; Biometra, Göttingen, Germany) in a 25-μl reaction mixture containing template DNA (5 μl of boiled lysate), 10 pmol of forward and reverse primers (Table ​(Table1),1), 0.2 mM deoxynucleoside triphosphate mixture, 0.625 U Taq DNA polymerase, and 2.5 mM MgCl₂ in 1× PCR buffer (Fermentas, Vilnius, Lithuania). The amplification schemes were based on previous amplification protocols (Table ​(Table1).1). For amplification of the tcpC sequence, the following amplification scheme was employed: 1 cycle of denaturation at 94°C for 4.5 min, followed by 25 cycles of denaturation at 94°C for 30 s, annealing at 60°C for 30 s, and elongation at 72°C for 1 min. The amplification was concluded with an extension program of one cycle at 72°C for 5 min. Fisher''s exact test (two-tailed; http://www.langsrud.com/fisher.htm) and the Bonferroni correction were used to analyze the data. The threshold for statistical significance after the Bonferroni correction was set at a P value of <0.05. The PCR revealed that 49 (23%) of the pathogenic strains studied harbored the tcpC sequence: 23 (21%) of our UTI E. coli isolates (21 isolates [21%] from individuals with symptomatic UTIs and 2 isolates [20%] from individuals with asymptomatic UTIs) and 26 (25%) of our SSTI E. coli isolates. The prevalence of tcpC was much lower among commensal E. coli isolates, only 7 (8%), as was found in a recent study by Cirl et al. (1). Comparison of the prevalence of tcpC among the UTI isolates of the two studies was not possible, as we could not obtain data on the type of symptomatic UTI (cystitis, pyelonephritis), and furthermore, the number of asymptomatic UTI isolates was too small (n = 10) to be statistically relevant. As seen from Table ​Table2,2, strong statistical correlations were found between the presence of tcpC and the B2 phylogenetic group, as well as between the presence of tcpC and the presence of cnf1, hlyA, papGIII, sfaDE, and usp among UTI isolates, as well as commensal strains. Among the SSTI isolates, statistically significant associations were found only between the presence of tcpC and the presence of cnf1, hlyA, and usp. As ExPEC strains mainly belong to the B2 phylogenetic group, these correlations and the higher virulence scores of the tcpC-encoding strains are not surprising. Interestingly, when the UTI and SSTI isolates were compared, major differences were observed. While the prevalence rates of tcpC sequences were similar in both groups, 21% among UTI isolates and 25% among SSTI isolates, suggesting an important role of TcpC in UTIs as well as in SSTIs, P values establishing significant correlations were higher among UTI isolates than among SSTI isolates. The differences between the UTI and SSTI E. coli strains observed are most likely due to differences in pathogenic mechanisms; nevertheless, the possession of TcpC seems to be an important factor in establishing UTIs and SSTIs. As the bowel flora is a reservoir of ExPEC, it is not surprising that tcpC was also found to be significantly associated with the B2 phylogenetic group among commensal strains. Our results suggest that even though E. coli strains able to induce disease outside the gastrointestinal tract are collectively designated ExPEC (11), it could be worthwhile to consider strains from different sites or syndrome-specific pathotypes separately.

TABLE 1.

Sequences of primers used in this study

转录因子FoxO3a在小鼠牙胚发育过程中的表达

目的：通过观察转录因子FoxO3a在小鼠牙胚发育中的表达特点及分布情况,探讨其在小鼠牙胚发育过程中的作用。方法取出生0 d和3 d的昆明小鼠各3只,处死解剖下颌骨,放于新鲜配置的10％中性甲醛溶液中固定过夜,制备小鼠牙胚0d和3d的组织标本,常规石蜡包埋,近远中向5μm连续切片,采用免疫组化方法观察检测FoxO3 a在小鼠不同时期的牙胚表达情况和分布情况,并对结果进行分析。结果 FoxO3 a在小鼠不同时期的牙胚发育中的分布存在差异,FoxO3a在0d小鼠牙胚中,成牙本质细胞成阴性表达,在3d小鼠牙胚中成牙本质细胞成阳性表达。结论观察FoxO3a在小鼠牙胚发育过程中的表达以及表达的变化,提示FoxO3a可能参与牙本质的形成和矿化。     

阻塞性睡眠呼吸暂停综合征与高脂血症关系长期随访研究

目的随访阻塞性睡眠呼吸暂停综合征(OSAS)与高脂血症的关系。方法对1868人进行前瞻性随访,通过家访式调查,随访期间每年进行1次体检,检查血压、血脂、血糖、心电图及X线胸片等,随访时间为20年。结果确诊OSAS者598/1868(32.0%),男496例(82.9%),女102例(17.1%);无OSAS者(对照组)1270/1868(68.0%)。随访终点OSAS组高脂血症529例,对照组396例(P<0.01)。结论 OSAS患者发生高脂血症可能性较一般人群高,考虑OSAS与高脂血症发生存在相关性。提示OSAS可能是高脂血症的独立危险因素。引起高脂血症的机制可能与OSAS导致食欲增加、活动减少、肥胖等有关。     

Novel Clostridium perfringens enterotoxin suicide gene therapy for selective treatment of claudin-3- and -4-overexpressing tumors

Bacterial toxins are known to be effective for cancer therapy. Clostridium perfringens enterotoxin (CPE) is produced by the bacterial Clostridium type A strain. The transmembrane proteins claudin-3 and -4, often overexpressed in numerous human epithelial tumors (for example, colon, breast, pancreas, prostate and ovarian), are the targeted receptors for CPE. CPE binding to them triggers formation of membrane pore complexes leading to rapid cell death. In this study, we aimed at selective tumor cell killing by CPE gene transfer. We generated expression vectors bearing the bacterial wild-type CPE cDNA (wtCPE) or translation-optimized CPE (optCPE) cDNA for in vitro and in vivo gene therapy of claudin-3- and -4-overexpressing tumors. The CPE expression analysis at messenger RNA and protein level revealed more efficient expression of optCPE compared with wtCPE. Expression of optCPE showed rapid cytotoxic activity, hightened by CPE release as bystander effect. Cytotoxicity of up to 100% was observed 72?h after gene transfer and is restricted to claudin-3-and -4-expressing tumor lines. MCF-7 and HCT116 cells with high claudin-4 expression showed dramatic sensitivity toward CPE toxicity. The claudin-negative melanoma line SKMel-5, however, was insensitive toward CPE gene transfer. The non-viral intratumoral in vivo gene transfer of optCPE led to reduced tumor growth in MCF-7 and HCT116 tumor-bearing mice compared with the vector-transfected control groups. This novel approach demonstrates that CPE gene transfer can be employed for a targeted suicide gene therapy of claudin-3- and -4-overexpressing tumors, leading to the rapid and efficient tumor cell killing in vitro and in vivo.