Multislice computerized tomography coronary angiography can be a comparable tool to intravascular ultrasound in evaluating "true" coronary artery bifurcations.

Aim: Coronary bifurcation atherosclerosis depends on its angles, flow, and extensive branching. We investigate the ability of CT coronary angiography (CTCA) to determine atherosclerotic plaque characteristics of "true" bifurcation compared with intravascular ultrasound (IVUS) and the influence on side branch (SB) fate after percutaneous coronary intervention (PCI). Methods and results: The study included 70 patients with 72 "true" bifurcations. Most of the bifurcations were in the left anterior descending-diagonal (Dg) territory [50 out of 72 (69.4%)]. Longitudinal plaque evaluation at the polygon of confluence [carina and 5 mm proximal and distal in the main branch (MB)] showed that carina side MB and SB plaque had occurred with the lowest incidence with fibro-lipid structure (115 ± 63 HU and 89 ± 73 HU, p < 0.001 for all). Bland-Altman analysis showed a discrepancy in measuring mainly the lumen area between CTCA and IVUS in proximal MB [lumen 5.10, 95% CI (95% confidence interval, 4.53-5.68) mm2, p < 0.001; vessel -1.42, 95% CI (-2.63 to -0.21) mm2, p = 0.023], carina MB [lumen 3.74, 95% CI (3.37-4.10) mm2, p < 0.001; vessel -0.48, 95% CI (-1.45 to 0.48) mm2, p = 0.322], and distal MB [lumen 4.72, 95% CI (4.27-5.18) mm2, p < 0.001; vessel 0.62, 95% CI (-0.53 to 1.77) mm2, p = 0.283]. A significant correlation existed between average plaque density on CTCA with a percentage of calcified plaque on IVUS tissue characterization (proximal r = 0.307/p = 0.024, carina 0.469/0.008, distal 0.339/0.024, minimal lumen diameter 0.318/0.020). Circumferential plaque in the proximal MB segment remained an independent predictor of SB compromise [OR 3.962 (95% CI 1.170-13.418)]. Conclusion: Detection and characterization of atherosclerotic plaque by CTCA in non-left main "true" coronary bifurcations can provide useful information about bifurcation anatomy and plaque distribution that can predict outcomes after provisional stenting, thus guiding the interventional strategy to bifurcation PCI.

Extracorporeal Life Support for Patients With Newly Diagnosed HIV and Acute Respiratory Distress Syndrome: A Systematic Review and Analysis of Individual Patient Data.

Extracorporeal membrane oxygenation (ECMO) may improve survival in patients with severe acute respiratory distress syndrome (ARDS). However, presence of immunosuppression is a relative contraindication for ECMO, which is withheld in HIV patients. We performed a systematic review to investigate the outcome of newly diagnosed HIV patients with ARDS receiving ECMO support. Our search yielded 288 publications, with 22 studies finally included. Initial presentation included fever, respiratory distress, and cough. Severe immunodeficiency was confirmed in most patients. Deceased patients had a higher viral load, a lower Horovitz index, and antiretroviral therapy utilized before ECMO. Moreover, ECMO duration was longer ( p = 0.0134), and all deceased suffered from sepsis ( p = 0.0191). Finally, despite the development of therapeutic options for HIV patients, ECMO remains a relative contraindication. We found that ECMO may successfully bridge the time for pulmonary recovery in 93% of patients, with a very good outcome. Using ECMO, the time for antimicrobial therapy, lung-protective ventilation, and immune system restitution may be gained. Further studies clarifying the role of ECMO in HIV are crucial and until these data are available, ECMO might be appropriate in immunocompromised patients. This holds especially true in newly diagnosed HIV patients, who are usually young, without comorbidities, with a good rehabilitation potential.

Acute Coronary Syndrome Presenting during On- and Off-Hours: Is There a Difference in a Tertiary Cardiovascular Center?

Background and Objectives: ACS presents an acute manifestation of coronary artery disease and its treatment is based on timely interventional diagnostics and PCI. It has been known that the treatment and the outcomes are not the same for all the patients with ACS during the working day, depending on the availability of the procedures and staff. The aim of the study was to explore the differences in clinical characteristics and outcomes in patients admitted for ACS during on- and off-hours. Materials and Methods: The retrospective study included 1873 consecutive ACS patients admitted to a tertiary, university hospital that underwent coronary angiography and intervention. On-hours were defined from Monday to Friday from 07:30 h to 14:30 h, while the rest was considered off-hours. Results: There were more males in the off-hours group (on-hours 475 (56%) vs. off-hours 635 (62%); p = 0.011), while previous MI was more frequent in the on-hours group (on 250 (30%) vs. off 148 (14%); p < 0.001). NSTEMI was more frequent during on-hours (on 164 (19%) vs. off 55 (5%); p < 0.001), while STEMI was more frequent during off-hours (on 585 (69%) vs. off 952 (93%); p < 0.001). Patients admitted during on-hours had more multivessel disease (MVD) (on 485 (57%) vs. off 489 (48%); p = 0.006), as well as multivessel PCI (on 187 (22%) vs. off 171 (16%); p = 0.002), while radial access was preferred in off-hours patients (on 692 (82%) vs. off 883 (86%); p = 0.004). Left main PCI was performed with similar frequency in both groups (on 37 (4%) vs. off 35 (3%); p = 0.203). Death occurred with similar frequency in both groups (on 17 (2.0%) vs. off 26 (2.54%); p = 0.404), while major adverse cardio-cerebral events (MACCEs) were more frequent in the on-hours group (on 105 (12.4%) vs. off 70 (6.8%); p = 0.039) probably due to the more frequent repeated PCI (on 49 (5.8%) vs. off 27 (2.6%); p = 0.035). Conclusions: Patients admitted for ACS during working hours in a tertiary hospital present with more complex CAD, have more demanding interventions, and experience more MACCEs during follow-up mostly due to myocardial infarctions and repeated procedures.

Idiopathic premature ventricular complexes treatment: Comparison of flecainide, propafenone, and sotalol.

BACKGROUND: Beta-blockers (BB) or dihydropyridine calcium channel blockers (CCBs) are still the first choices in the treatment of idiopathic premature ventricular complexes (PVCs), with low-modest efficacy. Antiarrhythmic drugs (AADs) of Ic class are moderate to highly efficient but the evidence on their benefits is still limited. AIM: To compare effectiveness and safety of flecainide, propafenone, and sotalol in the treatment of symptomatic idiopathic PVCs. METHODS: Our single-center retrospective study analyzed 104 consecutive patients with 130 medication episodes of frequent idiopathic PVCs treated with AADs flecainide, propafenone (Ic class) or sotalol (III class). The primary outcome was complete/near complete reduction of PVCs after medication episode (PVCs burden reduction >99%), and the secondary outcome was significant PVC burden reduction (≥80%). RESULTS: The complete/near complete PVCs burden reduction occurred in 31% and was significant in 43% of treated patients. A reduction of PVC burden for >99% was achieved in 56% of patients on flecainide, in 11% of patients on propafenone (p = .002), and in 21% of patients receiving sotalol (p = .031). There was no difference between propafenone and sotalol (p = .174). A reduction of PVC burden for ≥80% was achieved in 64% of patients on flecainide, in 30% of patients on propafenone (p = .009), and 33% of patients on sotalol (p = .020). There was no difference between propafenone and sotalol (p = .661). CONCLUSIONS: The efficacy of AADs class Ic and III in the treatment of idiopathic PVCs was modest. Flecainide was the most effective AAD in the achievement of complete/near complete or significant PVC burden reduction, compared to propafenone and sotalol.

Ablation of residual potentials along the circumferential line reduces acute pulmonary vein reconnection.

INTRODUCTION: Acute pulmonary vein (PV) reconnection is frequently encountered in patients undergoing PV isolation (PVI) procedure for the treatment of atrial fibrillation. In this study, we investigated whether the identification and ablation of residual potentials (RPs), after the initial achievement of PVI, reduces acute PV reconnection rate. METHODS: Following PVI in 160 patients, mapping along the ablation line was performed to identify RPs, defined as bipolar amplitude ≥0.2 mV or 0.1-0.19 mV combined with a negative component of the unipolar electrogram. Ipsilateral PV sets with RPs were randomized to either no further ablation (Group B) or to additional ablation of the identified RPs (Group C). The primary study endpoint was spontaneous or adenosine-mediated acute PV reconnection after a 30-min waiting period and was also evaluated in ipsilateral PV sets without RPs (Group A). RESULTS: After isolation of 287 PV pairs, 135 had no RPs (Group A), whereas the remaining PV pairs were randomized to either Group B (n = 75) or Group C (n = 77). Ablation of RPs resulted in a reduction of spontaneous or adenosine-mediated PV reconnection rate (16.9% in Group C vs 48.0% in Group B; p < 0.001). Group A was associated with a significantly lower percentage of acute PV reconnection as compared to Group B (5.9% vs 48.0%; p < 0.001) and Group C (5.9% vs 16.9%; p = 0.016). CONCLUSION: After PVI achievement, the absence of RPs along the circumferential line is associated with a low likelihood of acute PV reconnection rate. Ablation of RPs significantly reduces spontaneous or adenosine-mediated acute PV reconnection rate.

Pulmonary Vein Isolation Outcome Degree Is a New Score for Efficacy of Atrial Fibrillation Catheter Ablation.

This study introduces the pulmonary vein isolation outcome degree (PVIOD) as a new semiquantitative measure for the efficacy of atrial fibrillation (AF) catheter ablation and reports the determination of predictors associated with PVIOD. The median follow-up periods of 117 patients after the first and last ablation were, respectively, 82 (IQR 15) and 72 (IQR 30) months. PVIOD 1 included 32.5% of patients, those with successful single pulmonary vein isolation (PVI); PVIOD 2 included 29.1% of subjects, those with success after multiple procedures; PVIOD 3 comprised 14.5% of patients, those with clinical success; and PVIOD 4 included 23.9% of cases, those with procedural and clinical failure. In the multivariate ordinal logistic regression analysis, PVIOD 1-4 were independently associated with longstanding persistent AF with paroxysmal AF as the referent category (odds ratio (OR), 3.5; 95% confidence interval (95% CI), 1.1-10.7 (p = 0.031)), left atrial (LA) diameter (OR, 1.2; 95% CI, 1.1-1.3 (p = 0.001)) and left ventricular ejection fraction (LVEF) (OR, 0.9; 95% CI, 0.86-1.0 (p = 0.038)). LA size > 41 mm, LVEF ≤ 50% and longstanding persistent AF are strong predictors of AF recurrence. PVIOD 1-4 offer the most exact long-term prognosis of PVI. The purpose of the present article is to expand the quantitative measure of procedural success in the medical and biological fields.

Ablation of typical atrial flutter guided by the paced PR interval on the surface electrocardiogram: a proof of concept study.

AIMS: We aimed to assess the novel concept of using the paced PR interval (PRI) on the surface electrocardiogram (ECG) to prove trans-isthmus block after cavotricuspid isthmus (CTI) ablation for typical atrial flutter (AFl). METHODS AND RESULTS: Consecutive patients with AFl underwent linear radiofrequency ablation of the inferior CTI (6 o'clock). After AFl termination and/or presumed completion of the CTI line, CTI block was proven by atrial pacing by the ablation catheter medial (5 o'clock) and lateral to the line (7 and 9 o'clock). Corresponding PRIs were measured on the surface ECG. CTI block was assumed, if a sudden increase in the PRI was observed by moving the pacing site from 5 to 7 o'clock, and if the latter was longer than at 9 o'clock. Afterwards, bidirectional CTI block was confirmed by differential pacing. Thirty-one patients (mean age 67 ± 16 years, 81% male) underwent CTI ablation, and 18/31 (58%) were in AFl at the time of ablation (cycle length 249 ± 31 ms). Successful CTI block as defined by the PRI method was achieved in 31/31 (100%), and the mean PRIs during pacing at 5, 7, and 9 o'clock were 203 ± 56 ms, 329 ± 70 ms, and 296 ± 66 ms, respectively. Cavotricuspid isthmus block was confirmed in all patients (100%) by coronary sinus pacing with a reversal of the local activation sequence lateral to the isthmus line. CONCLUSION: The method of PRI analysis on the surface ECG to guide CTI ablation is easy to apply and highly accurate in confirming CTI block. This simple technique enables the novel concept of CTI ablation and proof of block with a single catheter.

A rare case of pacemaker lead endocarditis successfully treated with open heart surgery.

BACKGROUND: Cardiac device-related endocarditis has emerged as a serious complication in the era of advanced medical technology. Pacemaker related infections are rare and life-threatening with incidence from 0.06% to 7% and high mortality rate (30-35%). Diagnosis is hard, frequently delayed and could be even missed due to poor clinical findings. The average delay in diagnosis is 5.5 month. We report a case of the late-onset of pacemaker lead endocarditis caused by S. epidermidis successfully treated with open heart surgery. CASE REPORT: Patient with persistent high fever for 11 month and suspicion for infective endocarditis was admitted in Cardiovascular Institute. No clinical signs of endocarditis were observed. TTE revealed large vegetation 30 × 17 mm attached to the atrial electrodes with high embolic potential. This finding was verified by transesophageal echocardiography (TEE), although CT scan did not reveal vegetation. Blood cultures were negative. A sternotomy with cardiopulmonary bypass was performed and electrodes were extracted with large vegetation. Intraoperative finding revealed large thrombus with vegetation around pacemaker leads. Cultures of the electrodes and vegetation revealed Staphylococcus epidermidis. Surgery was followed up with antibiotic treatment for 6 weeks. He has been followed up for the next 2 years, and without complications. CONCLUSION: The absence of criteria for endocarditis and negative blood cultures should not keep the physician from ruling out lead endocarditis. This complication carries high risk of mortality if left untreated.

Impact of Voiding and Incontinence Symptoms on Health-Related Life Quality in Serbian Male Population.

PURPOSE: To investigate the impact of lower urinary tract symptoms on health-related quality of life (QoL) in Serbian population considering socio-demographic characteristics, habits, and health status. MATERIALS AND METHODS: The study was conducted in the Primary Healthcare Center "Novi Beograd", Serbia. The study included 1424 male participants, aged 40 years and above. QoL was assessed by using the -36Item Short Form Health Survey (SF36-) questionnaire, while voiding and incontinence symptoms were measured using the International Continence Society Male Short Form (ICS male SF) questionnaire. RESULTS: Voiding and incontinence symptoms significantly correlate with all domains of QoL. Voiding and incontinence symptoms have a high influence on general health, social functioning, physical functioning and body pain. After adjusting for age and education, voiding and incontinence symptoms had a similar influence on QoL. In the multivariate model the influence of cardiovascular diseases and income on QoL was lower than voiding and incontinence symptoms.. CONCLUSION: Voiding and incontinence symptoms affect QoL domains differently. Incontinence symptoms have a greater impact on QoL than voiding symptoms.

Comparative analysis of the current payment system for hospital services in Serbia and projected payments under diagnostic related groups system in urology.

BACKGROUND/AIM: Global budget per calendar year is a traditional method of funding hospitals in Serbia. Diagnose related groups (DGR) is a method of hospital payment based on classification of patients into groups with clinically similar problems and similar utilization of hospital resources. The aim of this study was to compare current methods of hospital services payment with the projected costs by DRG payment method in urology. METHODS: The data were obtained from the information system used in the Clinical Hospital Center "Dr. Dragisa Misovic"--Dedinje in Belgrade, Serbia. The implemented hospital information system was the main criterion for selection of healthcare institutions. The study included 994 randomly selected patients treated surgically and conservatively in 2012. RESULTS: Average costs under the current payment method were slightly higher than those projected by DRG, however, the variability was twice as high (54,111 ± 69,789 compared to 53,434 ± 32,509, p < 0.001) respectively. The univariate analysis showed that the highest correlation with the current payment method as well as with the projected one by DRG was observed in relation to the number of days of hospitalization (ρ = 0.842, p < 0.001, and ρ = 0.637, p < 0.001, respectively). Multivariate regression models confirmed the influence of the number of hospitalization days to costs under the current payment system (ß = 0.843, p < 0.001) as well as under the projected DRG payment system (ß = 0.737, p < 0.001). The same predictor was crucial for the difference in the current payment method and the pro- jected DRG payment methods (ß = 0.501, p < 0.001). CONCLUSION: Payment under the DRG system is administratively more complex because it requires detailed and standardized coding of diagnoses and procedures, as well as the information on the average consumption of resources (costs) per DRG. Given that aggregate costs of treatment under two hospital payment methods compared in the study are not significantly different, the focus on minor surgeries both under the current hospital payment method and under the introduced DRG system would be far more cost-effective for a hospital as great variations in treatment performance (reductions of days of hospitalization and complications), and consequently invoiced amounts would be reduced.

[Pulmonary embolism as a cause of cardiac arrest: hypothermia in post-resuscitation period (cooling therapy)].

INTRODUCTION: Pulmonary embolism as a possible cause of acute heart failure is a potentially fatal condition that can cause death in all age groups. Patients successfully resuscitated after cardiac arrest have a high risk of increased mortality and their poor long-term outcome is often associated with severe neurological complications. CASE OUTLINE: This is a case report of a 67-year-old man after a successful cardiopulmonary resuscitation (CPR) which was followed by therapeutic hypothermia (TH). The patient visited the dermatological outpatients' department with clinical presentation of pain and swelling of the right leg, shortness of breath and chest pain. During examination the patient lost consciousness, stopped breathing and had cardiac arrest. ECG was done which registered asystole. We began CPR. After 59 minutes of resuscitation return of heartbeat was achieved. The patient was transported to the Emergency Department. On admission, after computerized tomography (CT) of the chest confirmed massive pulmonary embolism (PE), the patient was administered thrombolytic therapy with Metalyse (tenecteplase) and anticoagulation therapy (heparin). After stabilization, therapeutic hypothermia was applied. Combination of EMCOOLSpad on the chest and abdomen and cold Ringer lactate 500 ml at 4 degrees C was flushed. Temperature was decreased to 33 degrees C and kept stabile for 24 hours. After eight days the patient was conscious with a minimal neurological deficit. CONCLUSION: As shown in this case report, and according to the rich experience elsewhere, cooling therapy after out-of-hospital cardiac arrest and successful CRP may be useful in preventing neurological complications.

[Brugada syndrome--a brand new case].

BACKGROUND: Brugada syndrome (BS) is a disorder characterized by syncope or sudden death associated with one of several electrocardiographic (ECG) patterns characterized by incomplete right bundle branch block and ST elevation in the anterior precordial leads. Patients with BS are prone to develop ventricular tachyarrhythmias that may lead to syncope, cardiac arrest, or sudden cardiac death. CASE REPORT: A 58-year-old woman is the first described case of Brugada syndrome in Serbia with intermittent typical changes in basic electrocardiography (ECG): ST segment elevation in the precordial chest leads like dome or coved--major form or type I. For the last 27 years the patient had suffered of palpitations and dizziness, without syncopal events. Her sister had died suddenly during the night in sleep. During 24-hour Holter monitoring the patient had ventricular premature beats during the night with R/T phenomenon and during the recovery phase of exercise testing had rare premature ventricular beats as the consequence of parasympatethic stimulation. Late potentials were positive. Echocardiography revealed left ventricular ejection fraction of 60%. We performed coronary angiography and epicardial coronary arteries were without significant stenosis and structural heart disease was excluded. In the bigining of the electrophysiological study ECG was normal, and after administration of Propaphenon i.v. Brugada syndrome unmasked with appearance of type I ECG pattern. A programed ventricular stimulation induced non sustained ventricular tachycardia. One-chamber implantable cardioverter defibrillator was implanted and the patient was treated with a combination od amiodarone and metoprolol per os. After one-year folow-up, there were no episodes of ventricular tachycardia and ventricular fibrillation. CONCLUSION: Brugada syndrome is a myocardial disorder which prognosis and therapy are related to presence of ventricular fibrillation or ventricular tachycardia. Electrophysiologicaly induced malignant ventricular disorders class I are indication for implantation of cardioverter defibrilator, as also occurred in presented patient.

Cloning, characterization and analysis by RNA interference of various genes of the Chelonus inanitus polydnavirus.

Successful parasitism of some endoparasitic wasps depends on an obligately symbiotic association with polydnaviruses. These unique viruses have a segmented genome consisting of circles of double-stranded (ds) DNA and do not replicate in the parasitized host. They are produced in the wasp's ovary and injected into the host along with the egg. Chelonus inanitus is an egg-larval parasitoid; its polydnavirus (CiV) has been shown to protect the parasitoid larva from the host's immune system and to induce developmental arrest in the prepupal stage. The genome of CiV consists of at least 10-12 segments and five have been sequenced up to now. Here, the complete (CiV12g2) or partial (CiV12g1, CiV16.8g1) cloning of three new CiV genes is reported. All three occur only on one viral segment and have no similarity to other known polydnavirus genes, with the exception of a high similarity of CiV12g1 to CiV14g1 and CiV12g2 to CiV14g2. Furthermore, the first attempt of in vivo application of RNA interference to study the function of polydnavirus genes is shown. Injection of dsRNA of two late- and one early- and late-expressed CiV genes into CiV/venom-containing host eggs partially rescued last-instar larvae from developmental arrest. Injection of the same dsRNAs into parasitized eggs partially reduced parasitoid survival, mainly by preventing the successful emergence of the parasitoid from the host. These viral genes thus seem to be involved in inducing developmental arrest and in keeping the cuticle soft, which appears to be necessary for parasitoid emergence and host feeding.

Stage-dependent expression of Chelonus inanitus polydnavirus genes in the host and the parasitoid.

Chelonus inanitus (Braconidae) is a solitary egg-larval parasitoid of Spodoptera littoralis (Noctuidae). Along with the egg it also injects polydnaviruses (CiV) and venom, which are prerequisites for successful parasitoid development. CiV protects the parasitoid from encapsulation by the host's immune system and induces a developmental arrest in the prepupal stage. The polydnavirus genome consists of several double-stranded circular DNA segments. Proviral DNA is integrated in the wasp's genome and virus replication is restricted to the wasp's ovary. Here, the analysis of eight CiV genes located on five different segments revealed four patterns of expression in the course of parasitization: early, late, persistent but variable, and early and late. The comparison between parasitized and CiV/venom only containing hosts indicated that the presence of the parasitoid larva modulates transcript levels. Haemocytes, fat body and nervous tissue contained viral transcripts, values being highest in haemocytes. Small amounts of CiV transcripts were also observed in parasitoid larvae and pupae, suggesting transcription from the proviral integrated form of viral DNA. This is the first comparative analysis of the expression patterns of several viral genes in both parasitized and CiV/venom only containing hosts over the entire period of parasitization, and it reveals intricate interactions between the parasitoid, the polydnavirus and the host.

Human tonsillar tissue block cultures differ from autologous tonsillar cell suspension cultures in lymphocyte subset activation and cytokine gene expression.

Lymphoid tissues cultured either as tissue blocks or as cell suspensions are used to study the behaviour of immune cells within their habitat. The preservation of tissue structures in tissue blocks, which is considered to be a major advantage, has been poorly defined. We characterised the morphological evolution of tissue cultures from human palatine tonsils and compared their lymphocyte subsets and the constitutive cytokine gene expression to those in autologous tonsillar single-cell suspension cultures over time, and after adding cyclosporin A (CsA) to mimic the situation in individuals treated with immunosuppressive drugs. Density and morphology of follicles were conserved up to 4 days, during which tissue cultures exhibited similar cell viability as suspension cultures, but a significantly less frequent increase of CD95 expression in T cells, smaller variation of the proportion of CD4(+) cells and better CD21(+)/CD23(-) B-cell survival. Treatment with cyclosporin A at higher concentrations resulted in superior histologic preservation of lymphoid tissue structures and seemed to further prevent the expression of CD95 by CD3(+) cells and the activation in tissue culture of CD21(+) cells. Constitutive gene expression levels of the stromal cytokines interleukin (IL)-1beta and interleukin-6 in tissue culture were significantly higher than those in suspension cultures. These results suggest that tonsillar tissue cultures preserve their structure only for a limited time, during which they more closely reflect processes in vivo, including a state of iatrogenic immunosuppression, than do their cell suspension counterparts.

Quantitative cytokine gene expression in human tonsils at excision and during histoculture assessed by standardized and calibrated real-time PCR and novel data processing.

Real-time reverse transcription polymerase chain reaction (RT-PCR) assays were developed for the quantification of expression of the genes for human interleukin (IL)-1beta, IL-2, IL-6, IL-8, IL-10, IL-12, IL-15, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta and for the endogenous reference hydroxymethylbilane synthase (HMBS). The assays detected as little as five plasmid copies and were 100% specific. The creation and integration of a calibration sample into the assays permitted their calibration across experiments. To handle the high number of generated data, the correlator of advanced real-time assays (CARTA) software was designed to organize samples and to automatically control and analyze TaqMan real-time RT-PCR data. The RT-PCR assays were applied to quantify levels of cytokine gene expression in human palatine tonsils at excision and during 4 days of histoculture. Similar longitudinal patterns of cytokine gene expression were observed in all donors, but the variations in spontaneous expression levels between donors were large. The expression levels in histocultures were constant over time and similar to the expression levels at excision except for IL-6 and IL-8, which markedly increased following the first 24 h of culture, possibly due to the initial stress. The standardized and calibrated RT-PCR assays quantify gene expression of human cytokines proved sensitive and specific for the investigation of cell behavior at the molecular level and the newly established CARTA software, a reliable tool for rapid data handling. Tonsil histocultures could serve as a valuable ex vivo model system for further, donor-dependent, studies on activation or repression of cytokine gene expression.