Detection of virulence genes of Clostridium difficile by multiplex PCR

Antikainen J, Pasanen T, Mero S, Tarkka E, Kirveskari J, Kotila S, Mentula S, Könönen E, Virolainen‐Julkunen A‐R, Vaara M, Tissari P. Detection of virulence genes of Clostridium difficile by multiplex PCR. APMIS 2009; 117: 607–13. Clostridium difficile strains belonging to the PCR ribotype 027, pulse‐field gel electrophoresis (PFGE) type NAP1, toxinotype III and restriction endonuclease analysis group BI harbouring mutations in the tcdC gene and possessing binary toxin components A and B have been described to cause epidemics with increased morbidity and mortality. In the present study we developed a conventional multiplex PCR designed to detect selected virulence associated markers of the hypervirulent C. difficile PCR ribotype 027. The multiplex PCR assay detected the major toxins A and B, binary toxin components A and B as well as a possible deletion in the tcdC gene: a characteristic pattern of amplification products for the PCR ribotype 027 strains was detected. This rather simple method was specific for the screening of this hypervirulent C. difficile strain. The correlation between the multiplex PCR and PCR ribotyping methods was excellent. The sensitivity and specificity were 100% in our epidemiological situation. In conclusion, this multiplex PCR was found useful in the preliminary screening for the hypervirulent C. difficile PCR ribotype 027.     

Enhancement of Tuberculin-Induced Lymphocyte Transformation by Precultivated Macrophages from Patients with Sarcoidosis

Precultivated autologous macrophages enhanced the early PPD-induced blastogenic response of tuberculin-sensitive lymphocytes. This effect was not caused by a diffusible macrophage product. Macrophages exposed to PPD and subsequently washed caused as high a response as did PPD in cultures without precultivated macrophages. Sensitive macrophages were not able to induce nonsensitive lymphocytes to respond to PPD. Precultivated macrophages from tuberculin-negative patients with sarcoidosis enhanced the PPD-induced transformation of sensitive lymphocytes, as did autologous macrophages Cord blood-derived macrophages were significantly less effective     

An outbreak of pneumonia associated with S. pneumoniae at a military training facility in Finland in 2006

Streptococcus pneumoniae is a well‐known cause of community‐acquired bacterial pneumonia. The purpose of this study was to assess the cause and extent of the outbreak of pneumonia which occurred among military recruits following a 1‐week hard encampment in Finland. We also assessed the carriage rate and molecular characteristics of the S. pneumoniae isolates. All pneumococcal isolates were studied for antibiotic susceptibility, serotyped, genotyped by multilocus sequence typing (MLST), and the presence of pneumococcal rlrA pilus islet was detected. The genotype results defined by MLST corresponded with the serotype results. S. pneumoniae serotype 7F, ST2331, seemed to be associated with an outbreak of pneumonia and nasopharyngeal carriage among 43 military recruits. Of the 43 military recruits, five (12%) were hospitalized with pneumonia and two (40%) of them were positive for S. pneumoniae serotype 7F, ST2331 by blood culture. Eighteen (42%) of the 43 men were found to be positive for S. pneumoniae by nasopharyngeal culture, and nine (50%) of them carried pneumococcal serotype 7F, ST2331. The outbreak strain covered 55% of all the pneumococcal findings. Outbreaks of invasive pneumococcal disease seem to occur in a crowded environment such as a military training facility even among previously healthy young men.     

Is the Expression of Classical HLA Class I Antigens on Trophoblast of Importance for Human Pregnancy?

PROBLEM: Human leukocyte antigen (HLA)-C and possibly also HLA-B seem to be expressed on the extravillous trophoblast. These antigens carry epitopes that function as ligands for natural killer (NK)-cell-inhibitory receptors. Antitrophoblast cytotoxicity mediated by decidual NK cells might be involved in miscarriage. We thus found it relevant to elucidate whether parental HLA-C and -Bw polymorphism play a role in recurrent miscarriage (RM). METHOD OF STUDY: HLA-C and -Bw investigations by DNA-based techniques were undertaken in 35 couples with unexplained RM and in 30 couples with normal fecundity. The number of HLA-C- and -Bw-related supertypic specificities that can bind NK-cell-inhibitory receptors was evaluated in selected couples. RESULTS: The proportions of couples with RM and control couples carrying four HLA-C alleles with the same NK-cell-inhibitory supertypic specificities were equal. In 46% of studied couples with RM, all four HLA-B alleles carried the HLA-Bw6 supertypic specificity, which was significantly higher than the corresponding frequency (17%) in the control couples (P < 0.02). CONCLUSIONS: The expression of polymorphic HLA-C on trophoblasts does not seem to play a role in RM. Assuming that HLA-B is expressed on trophoblasts, we may suggest that the revealed predominance of HLA-Bw6 expression (which excludes the presence of HLA-Bw4—protective antigens) may predispose a particular couple to the RM phenomenon.     

Outcome of Alcoholism: the structure of patient attributions as to what causes change

Various strands of the research literature which have bearing on the concept of attribution for change in drinking behaviour are reviewed. The reasons which subjects give for amelioration in their drinking are likely to reflect a mixture of the way in which change is being subjectively construed together with report on the influences and processes which may objectively contribute to change. We present data on such attributions as given by 66 alcoholics on whom we have detailed 10-year information on multiple aspects of outcome and who also completed a 70-item Attributions Inventory. A factor analysis of the Attributions data reveals two factors which have been labelled respectively ‘Active’ and ‘Responsive’, and it is suggested that this may be a more appropriate analytical framework than the single dimension of Internal/External locus of control. The relationship between Attribution factors on the one hand and Outcome components and clusters on the other, is explored.‘Active’ Attribution is significantly and positively related to better ‘High Dependence’ Outcome, whereas ‘Responsive’ Attribution relates significantly and positively to better ‘Low Dependence’ outcome. Potential theoretical and clinical implications are discussed.     

Evolution of heart rate variability in cardiac transplant recipients: a clinical study

Koskinen P, Virolainen J, Koskinen Pk, Häyry P, Kupari M (Division of Cardiology, Department of Medicine, Helsinki University Central Hospital, and Transplantation Laboratory, University of Helsinki, Helsinki, Finland). Evolution of heart rate variability in cardiac transplant recipients: a clinical study. J Intern Med 1996; 239 : 443–9. Objectives. To investigate in cardiac transplant patients whether post-transplantation time, graft arteriosclerosis, allograft rejection, or earlier cytomegalovirus infection affect the neural regulatory mechanisms of the donor heart. Design. A consecutive series of heart transplant patients during a 12-month period. Setting. A university hospital in Finland. Subjects. Consecutive cardiac transplant recipients (n=38) attending the hospital for their annual clinical examintaion were studied. Their mean (SD) age was 45.4 (11.5) years, 37 were male, and the median (range) time since transplantation was 36 (12–72) months. Interventions. Power spectral analysis of R–R intervals (during 5 min of controlled breathing, the Valsalva manoeuvre, and deep breathing), routine coronary arteriography, cytomegalovirus serology. Results. R–R interval (r=0.67; P<0.001), the root mean square difference of successive R–R intervals (r=0.38; P<0.05), the total R–R interval power (r=0.45; P<0.01), the power of the very low frequency (0.0–0.07 Hz) component (r=0.53; P<0.01), and the power of the nonrespiratory (0.0–0.15 Hz) component (r=0.49; P<0.01) were related to the length of time since the operation. Patients having had a transplantation 3 years ago or more had significantly greater median (range) total R–R interval power than those having had the operation less than 3 years ago (59 [10–265] vs. 20 [3–113] ms²; P=0.02). There was also a difference between the two groups in the very low frequency component (18 [1–226] vs. 5 [0–45] ms²; P=0.01), in the nonrespiratory component (30 [1–227] vs. 9 [0–53] ms²; P=0.02), and in the Valsalva ratio (0.995 [0.955–1.065] vs. 1.020 [0.975–1.155]; P=0.03). Patients with and without graft arteriosclerosis, episodes of rejection, or earlier cytomegalovirus infection showed no difference in the power spectral measures. Conclusions. The donor heart rate variability increases with post-transplantation time. Heart rate variability in transplant recipients is not related to the extent of graft arteriosclerosis, episodes of allograft rejection, or earlier cytomegalovirus infection.     

Response of subcutaneous and cutaneous metastases of malignant melanoma to combined cytostatic plus interferon therapy

Summary A chemotherapy regimen consisting of dacarbazine (DTIC), vincristine, bleomycin and lomustine (CCNU) was combined with natural leucocyte interferon (IFN) in the treatment of 37 patients with cutaneous and subcutaneous metastases of malignant melanoma. Twenty-five also had concomitant lymph node, visceral, bone or brain metastases. Fifteen patients (41%) experienced complete response (CR) and 10 (27%) partial response (PR) of the superficial lesions. In addition, three patients were rendered surgically tumour-free after PR or disease stabilization during drug therapy. The median overall duration of response was 10.2 months (range 1–53 months). In disseminated disease, the combined therapy produced favourable results, particularly with regard to superficial lesions. It is also possible that this therapy may retard the growth of aggressive subcutaneous metastases in patients who have previously had multiple surgical procedures in an attempt to stabilize their disease. In a small proportion of patients, the long-term complete remission with the drug therapy alone, or in combination with surgery, suggests that this regimen might have been curative.     

Influence of negative intrathoracic pressure on right atrial and systemic venous dynamics

To investigate the effect of a sustained fall in intrathoracicpressure (Mueller manoeuvre) on blood flow through the rightheart and on systemic venous dynamics, 16 patients were studiedusing thermodilution, cinevenograms and simultaneous pressurerecordings with two micromanometric transducers. The reductionsin airway pressure (median [rangeI) during two graded Muellermanoeuvres were 25 (20–30) and 42 (22–52) mmHg.Right atrial mean pressure decreased by 17 (2–25) mmHgduring the former and 38 (0–49) mmHg during the latter,and simultaneously, pressure gradients of 23 (1–32) and45 (1–82) mmHg developed between the inferior vena cavaand right atrium (P<0.003 for all). Internal jugular venouspressure decreased by 16 (4–25) and 24 (4–43) mmHg(P<0.03 for both), respectively, and no pressure gradientdeveloped between internal jugular and superior caval veins.The minimum diameter of the proximal inferior vena cava decreasedby 69 (–49–84)% (P = 0.002) during the greater manoeuvre.Cardiac index tended to increase by 26 (–17–40)%(P<0.066) during the lesser manoeuvre but did not changestatistically significantly during the greater. In conclusion,during negative intrathoracic pressure caused acutely by theMueller manoeuvre, right atrial pressure decreases and the inferiorvena cava collapses partially at or below the diaphragm. Despitea significant venous obstruction between the lower body andright atrium, blood flow through the right heart increases orremains constant.     

西藏高原地区腰麻联合硬膜外麻醉与硬膜外麻醉的临床比较研究

目的:探讨腰麻联合硬膜外阻滞在西藏高原地区用于下腹及下肢的手术的临床效果。方法:36例在腰麻联合硬膜外阻滞下行下肢及下腹部手术病人(CSEA组),根据手术需要选择单点法或双点法,以26 G腰麻穿刺针通过硬膜外针刺入蛛网膜下腔,注入局麻药物重比重0．5％布比卡因0．2 mg体重(Kg),记录两组病人入手术室时和麻醉后10、30、60 min 的血压、心率以及氧饱和度;麻醉到手术开始的时间;麻醉效果;术后恢复情况,有无并发症等。选择既往2004年硬膜外麻醉病例36例作为对照组(EA组)。结果:EA组在麻醉时间和手术开始时间均明显大于CSEA组(P<0．05);在麻醉后 10min,EA组舒张压明显下降(P<0．05),CSEA组收缩压及舒张压均明显下降,麻醉后30及60min,两组病人血压均有升高,与麻醉前对比无明显差异(P>0．05);EA组病人在麻醉后吸氧(2～4)L／min,麻醉后10、30、60 min SpO2都明显高于麻醉前(P<0．01);CSEA组麻醉后10 min在未吸氧的情况下SpO2明显高于麻醉前(P<0．05),麻醉后30、60 min吸氧(2～ 4)L／min SpO2明显高于麻醉前(P<0．01)。结论:CSEA在高原环境下是安全可行的,在下腹和下肢手术中是一个比较理想的麻醉选择,与硬膜外阻滞相比,其麻醉药用量小,作用快、效果确切,对呼吸影响轻微,但对有明显心肌缺血的病人应慎用。