Diagnostic usefulness of bone marrow aspiration material for the amplification of IS6110 insertion element in extrapulmonary tuberculosis: comparison of two PCR techniques.

SETTING: In many cases of extra-pulmonary tuberculosis (EPTB), with the exception of paucibacillary analysed specimens, the suspected site of mycobacterial infection is relatively inaccessible or unknown, making laboratory confirmation of TB laborious and problematic. OBJECTIVE: Two different polymerase chain reaction (PCR) based methods were compared to investigate the validity of bone marrow aspiration material as an easily accessible alternative sample for molecular analysis in EPTB. DESIGN: We amplified the same sequence of IS6110 of Mycobacterium tuberculosis complex in 19 confirmed cases of EPTB using two different nested PCR techniques: one in-house 'classic' PCR and another based on LightCycler technology. RESULTS: Both methods demonstrated the same reliability when performed in samples of infected tissue. However, the LightCycler protocol was superior to the in-house system when applied in bone marrow aspiration material, revealing positivity in 18/19 compared to 13/19 samples of 'classic' PCR. CONCLUSION: The application of an optimised LightCycler nested amplification protocol in bone marrow aspirates may promote diagnostic accuracy in difficult and/or urgent cases of EPTB.     

Comparison of standard external CPR, open-chest CPR, and cardiopulmonary bypass in a canine myocardial infarct model

STUDY OBJECTIVES: After cardiac arrest, open-chest CPR (OCCPR) and cardiopulmonary bypass (CPB) have demonstrated higher resuscitation rates when compared individually with standard external CPR (SECPR). We compared all three techniques in a canine myocardial infarct ventricular fibrillation model. TYPE OF PARTICIPANTS: Twenty-six mongrel dogs were block-randomized to receive SECPR and advanced life support (nine), CPB (nine), or OCCPR (eight). DESIGN AND INTERVENTIONS: All dogs received left anterior descending coronary artery occlusion followed by four minutes of ventricular fibrillation without CPR and eight minutes of Thumper CPR. At 12 minutes, dogs received one of three resuscitation techniques. After resuscitation, all animals received four hours of intensive care. Animals that were resuscitated had histochemical determination of ischemic and necrotic myocardial areas. MEASUREMENTS: Intravascular pressures were measured and coronary perfusion pressure was calculated during baseline, cardiac arrest, resuscitation, and postresuscitation periods. Percent necrotic myocardium, percent ischemic myocardium, and necrotic-to-ischemic ratios were determined for resuscitated animals. Epinephrine dosage and number of countershocks were determined for each group. MAIN RESULTS: Nine of nine CPB and six of nine OCCPR, compared with two of eight SECPR animals, were resuscitated (P less than .01). Three of nine CPB and OCCPR and two of eight SECPR dogs survived to four hours (P = NS). Coronary perfusion pressure two minutes after institution of technique was significantly higher with CPB (75 +/- 37 mm Hg) and OCCPR (56 +/- 31 mm Hg) than in SECPR animals (16 +/- 16 mm Hg, P less than .04). Epinephrine required for resuscitation was significantly less with CPB (0.10 +/- 0.02 mg/kg) than for SECPR (0.28 +/- 0.11 mg/kg, P less than .002). The ratio of necrotic to ischemic myocardium at four hours was significantly lower with CPB (0.15 +/- 0.31) and OCCPR (0.39 +/- 0.25) than for SECPR (1.16 +/- 0.31, P less than .02). CONCLUSION: OCCPR and CPB produce higher coronary perfusion pressures and improved resuscitation rates from ventricular fibrillation when compared with SECPR in this canine myocardial infarct cardiac arrest model. CPB and OCCPR yielded similar resuscitation results, although less epinephrine was required with CPB.     

Human herpesvirus 8 (HHV-8) infection in healthy urban employees from Greece: seroprevalence and associated factors

A cross-sectional study was carried out in healthy company employees from Greece with the aim of assessing the prevalence of human herpesvirus 8 (HHV-8) and identifying risk factors for this herpesviral infection. Serum samples obtained from 955 subjects were tested for antibodies to HHV-8 by the K8.1 enzyme-linked immunosorbent assay (ELISA). Associations between HHV-8 serostatus and potential risk factors were examined using t-test, chi square test, and multivariate logistic regression analysis. HHV-8 prevalence was 7.6% (95% confidence interval (CI): 6.0%, 9.5%) and it increased with age from 6.5% among <30 years old to 13.8% among > or =50 years old subjects (P = 0.006). HHV-8 seropositivity was independently associated with endoscopic examination (odds ratio (OR): 2.01; 95% CI: 1.09, 3.70; P = 0.026), HBsAg positivity (OR: 5.16; 95% CI: 2.02, 13.20; P = 0.001) and age (OR > or =50 years old vs. <50 years old: 2.09; 95% CI: 1.23, 3.52; P = 0.006). No statistically significant associations between HHV-8 positive status and gender, occupational status, surgery, transfusion, tattoos/body piercing, multiple sex partners, weakness/fatigue, HCV status were observed. HHV-8 is prevalent in Greece. The strong association between HBV infection and HHV-8 positive status supports the hypothesis of an association between these two viral infections. The association between HHV-8 seropositivity and endoscopic examination requires further investigation.     

Role of oxygen in postischemic myocardial injury

Myocardial function is dependent on a constant supply of oxygen from the coronary circulation. A reduction of oxygen supply due to coronary obstruction results in myocardial ischemia, which leads to cardiac dysfunction. Reperfusion of the ischemic myocardium is required for tissue survival. Thrombolytic therapy, coronary artery bypass surgery and coronary angioplasty are some of the treatments available for the restoration of blood flow to the ischemic myocardium. However, the restoration of blood flow may also lead to reperfusion injury, resulting in myocyte death. Thus, any imbalance between oxygen supply and metabolic demand leads to functional, metabolic, morphologic, and electrophysiologic alterations, causing cell death. Myocardial ischemia reperfusion (IR) injury is a multifactorial process that is mediated by oxygen free radicals, neutrophil activation and infiltration, calcium overload, and apoptosis. Controlled reperfusion of the ischemic myocardium has been advocated to prevent the IR injury. Studies have shown that reperfusion injury and postischemic cardiac function are related to the quantity and delivery of oxygen during reperfusion. Substantial evidence suggests that controlled reoxygenation may ameliorate postischemic organ dysfunction. In this review, we discuss the role of oxygenation during reperfusion and subsequent biochemical and pathologic alterations in reperfused myocardium and recovery of heart function.     

Compared to angiotensin II, epinephrine is associated with high myocardial blood flow following return of spontaneous circulation after cardiac arrest

INTRODUCTION: Epinephrine (adrenaline) and vasopressin are used currently to improve myocardial blood flow (MBF) during cardiac arrest. Angiotensin II has also been shown to improve MBF during CPR. We explored the effects of angiotensin II or epinephrine alone, and the combination of angiotensin with epinephrine, on myocardial and cerebral blood flows in a swine model of cardiac arrest. METHODS: Swine were instrumented for regional blood flow measurements. Ventricular fibrillation was induced and CPR begun. Angiotensin II 50 mcg/kg (ANG), epinephrine 0.02 mg/kg (EPI) or the combination (ANG+EPI) was administered. Blood flow was measured during baseline normal sinus rhythm (NSR), before (CPR) and after drug administration (CPR+DRUG), and post reperfusion return of spontaneous circulation (ROSC). RESULTS: All groups had a significant increase in MBF during CPR following drug administration (P<0.05). [table: see text] There was a trend toward higher flows in the EPI groups. The group receiving both EPI and ANG did not have higher blood flows than the EPI or ANG alone groups. Both groups that received EPI had markedly elevated MBF following ROSC compared with angiotensin II (P<0.05). CONCLUSIONS: The combination of ANG and EPI did not improve MBF during cardiac arrest. Epinephrine may increase MBF compared with angiotensin II post-reperfusion.     

Serum osmolal gap in clinical practice: usefulness and limitations

Background: Although serum osmolal gap can be a useful diagnostic tool, clinicians are not familiar with its use in clinical practice.

Objectives: The review presents in a series of questions-answers and under a clinical point of view the current data regarding the use of osmolal gap.

Discussion: The definition and the best formula used for the calculation of osmolal gap, the main causes of increased osmolal gap with or without increased anion gap metabolic acidosis, as well as the role of concurrent lactic acidosis or ketoacidosis are presented under a clinical point of view.

Conclusions: The calculation of osmolal gap is crucial in the differential diagnosis of many patients presenting in emergency departments with possible drug or substance overdose as well as in comatose hospitalized patients.     

Clinicopathological Assessment of Hepatitis C Virus Infection in Parenteral Drug Abusers

Objectives: To determine the severity of hepatic histological lesions in anti-HCV positive parenteral drug abusers and to correlate it with the level of ALT activity and HCV RNA determined by polymerase chain reaction (PCR).
Methods: Twenty-nine of the 62 anti-HCV-positive parenteral drug abusers who consecutively entered a Rehabilitation Center of Athens consented to liver biopsy and were prospectively and thoroughly followed up for a mean of 12.9 (range 6-33) months. Anti-HCV was detected by a second-generation enzyme immunoassay and confirmed by a second-generation recombinant immunoblot assay. Serum samples were tested for HCV RNA by nested PCR with primers from the highly conserved 5' untranslated region of the HCV genome.
Results: Liver biopsy revealed lesions compatible with chronic hepatitis in 26 (89.6%) and a normal liver in three (10.4%) of the 29 patients. In particular, 11 (37.9%) had minimal and 15 (57.1%) had mild chronic hepatitis; fibrosis was absent or mild in all cases. Histological grade and stage were significantly milder in patients with persistently normal ALT activity than in those with increased ALT activity. However, chronic hepatitis was observed in five (62.5%) of the eight patients with normal ALT levels. The presence of serum HCV RNA was not significantly correlated with the severity of histological lesions. HCV RNA was detected in 16 (57.1%) of the 28 cases tested. In particular, HCV RNA was detected in one (33.3%) of the three cases with normal liver and in three (37.5%) of the eight patients with normal ALT levels.
Conclusions: Liver biopsy appears to be the method of choice for the accurate evaluation of anti-HCV positive parenteral drug abusers, irrespective of ALT activity and presence of serum HCV RNA. Chronic hepatitis is observed in the majority and the state of "healthy" carrier of HCV in the minority of this epidemiological setting.