Paroxysmal atrial fibrillation occurs often in cryptogenic ischaemic stroke. Final results from the SURPRISE study.

BACKGROUND AND PURPOSE: Atrial fibrillation (AF) increases the risk of stroke fourfold and is associated with a poor clinical outcome. Despite work-up in compliance with guidelines, up to one-third of patients have cryptogenic stroke (CS). The prevalence of asymptomatic paroxysmal atrial fibrillation (PAF) in CS remains unknown. The SURPRISE project aimed at determining this rate using long-term cardiac monitoring. METHODS: Patients with CS after protocolled work-up including electrocardiography (ECG) and telemetry were included after informed consent. An implantable loop recorder (ILR) was implanted subcutaneously. PAF was defined by events of atrial arrhythmia >2 min with a correlating one-lead ECG confirming the diagnosis. RESULTS: Eighty-five patients were monitored for a mean of 569 days (SD ±310). PAF was documented in 18 patients (20.7%) during the study period and detected by ILR in 14 patients (16.1%). In three patients PAF was detected by other methods before or after monitoring and was undiscovered due to device sensitivity in one case. The first event of PAF was documented at a mean of 109 days (SD ±48) after stroke onset. PAF was asymptomatic in all cases and occurred in episodes lasting predominantly between 1 and 4 h. Four recurrent strokes were observed, three in patients with PAF; all three patients were on oral anticoagulation (OAC). CONCLUSIONS: One in five patients with CS had PAF, which occurred at low burden and long after stroke. Future studies should determine the role of implantable cardiac monitors after stroke and determine the potential therapeutic benefit of OAC treatment of patients with PAF.

High-risk human papillomavirus (hrHPV) E6/E7 mRNA testing by PreTect HPV-Proofer for detection of cervical high-grade intraepithelial neoplasia and cancer among hrHPV DNA-positive women with normal cytology.

Our aim was to investigate whether high-risk HPV (hrHPV) mRNA detection by PreTect HPV-Proofer can be used to stratify hrHPV DNA-positive women of different cytology classes for risk of high-grade cervical intraepithelial neoplasia or worse (cervical precancer or cancer, i.e., cervical intraepithelial neoplasia grade 2 or higher [≥ CIN2]). A total of 375 women participating in population-based screening, with a GP5+/6+-PCR hrHPV DNA-positive cervical scrape with normal cytology (n = 202), borderline or mild dyskaryosis (BMD) (n = 88), or moderate dyskaryosis or worse (>BMD) (n = 85), were enrolled. Cervical scrapes were additionally subjected to HPV16/18/31/33/45 E6/E7 mRNA analysis by PreTect HPV-Proofer (mRNA test). Referral and follow-up policies were based on cytology, hrHPV DNA, and mRNA testing. The primary study endpoint was the number of ≥CIN2 detected within 3 years of follow-up. The mRNA positivity increased with the severity of cytological abnormality, ranging from 32% (64/202) in hrHPV DNA-positive women with normal cytology to 47% (41/88) in BMD and 68% (58/85) in >BMD groups (P < 0.01). Women with ≥ CIN2 were more likely to test positive by mRNA test (63%) than women without evidence of ≥ CIN2 (32%; P < 0.01). A positive mRNA test result conferred an increased ≥ CIN2 risk in hrHPV DNA-positive women with normal cytology, i.e., 0.55 (95% confidence interval [95% CI], 0.34 to 0.76) in mRNA-positive versus 0.20 (95% CI, 0.07 to 0.33) in mRNA-negative women. In hrHPV DNA-positive women with BMD or >BMD, the result of the mRNA test did not influence the ≥ CIN2 risk. In conclusion, mRNA testing by PreTect HPV-Proofer might be of value to select hrHPV DNA-positive women with normal cytology in need of immediate referral for colposcopy.

A comprehensive evaluation of the accuracy of cervical pre-cancer detection methods in a high-risk area in East Congo.

BACKGROUND: Given the high burden of cervical cancer in low-income settings, there is a need for a convenient and affordable method for detecting and treating pre-cancerous lesions. METHODS: Samples for comparing the accuracy of cytology, virology and histology were collected. Identification of HPV E6/E7 mRNA was performed using PreTect HPV-Proofer. HPV DNA detection was performed by GP5+/6+ PCR, followed by reverse line blot (RLB) for typing. RESULTS: A total of 343 women, aged 25-60 years, attending gynaecological polyclinics in DR Congo were included for sample enrolment. The test positivity rate was conventional and liquid-based cytology (LBC) at cutoff ASCUS+ of 6.9 and 6.6%, respectively; PreTect HPV-Proofer of 7.3%; and consensus DNA PCR for 14 HR types of 18.5%. Sixteen cases of CIN2+ lesions were identified. Of these, conventional cytology identified 66.7% with a specificity of 96.2%, LBC identified 73.3% with a specificity of 96.9%, all at cutoff ASCUS+. HR-HPV DNA detected all CIN2+ cases with a specificity of 85.9%, whereas PreTect HPV-Proofer gave a sensitivity of 81.3% and a specificity of 96.6%. CONCLUSION: Both HPV detection assays showed a higher sensitivity for CIN2+ than did cytological methods. Detecting E6/E7 mRNA from only a subset of HR HPVs, as is the case with PreTect HPV-Proofer, resulted in a similar specificity to cytology and a significantly higher specificity than consensus HR HPV DNA (P<0.0001).

Dynamic autoregulation and renal injury in Dahl rats.

The Dahl salt-sensitive (Dahl S) rat develops hypertension and renal injuries when challenged with a high salt diet and has been considered to be a model of chronic renal failure. Renal injuries appear very early in life compared with the spontaneously hypertensive rat (SHR). During the course of hypertension, a gradual impairment of autoregulatory control of renal blood flow might expose the glomerular circulation to periods of elevated pressure, resulting in renal injuries in Dahl S rats. Dynamic autoregulatory capacity was assessed in Dahl S and Dahl salt-resistant (Dahl R) rats, SHR, and Sprague-Dawley rats by inducing broad-band fluctuations in the arterial blood pressure and simultaneously measuring renal blood flow. Dynamic autoregulation was estimated by the transfer function using blood pressure as the input and renal blood flow as the output. Renal morphological injuries were evaluated in Dahl S rats and SHR and were scored semiquantitatively. Dynamic autoregulation was efficient and comparable in the low-frequency range (<0.015 Hz) in Dahl R rats, SHR, and Sprague-Dawley rats. The response in Dahl S rats depended strongly on the initiation time of the high salt diet. Autoregulation was preserved during a low salt diet and in rats exposed to a late-onset hypertension of short duration, only partly preserved if the late-onset hypertension was of a longer duration, and abolished in early-onset hypertension. All Dahl S rats on a high salt diet showed severe morphological changes in the kidney. In conclusion, autoregulatory capacity in the kidney of Dahl S rats is gradually impaired when rats are rendered hypertensive with a high salt diet. Renal morphological injuries develop before loss of dynamic autoregulation. Impaired autoregulation appears to be the result, not the cause, of the process that ultimately leads to renal failure in the Dahl S rat.

Glicentin is present in the pig pancreas.

Specimens from porcine pancreas and ileal mucosa were extracted in acid/ethanol, subjected to gel permeation chromatography, ion-exchange chromatography, enzymatic peptide degradation, reverse-phase HPLC, and analysed for glucagon-like and glicentin-like immunoreactivity by region-specific radioimmunoassays. Results obtained with all methods were consistent with the hypothesis that glicentin is present in the pig pancreas in small amounts.

Human papilloma virus has no prognostic significance in cervical carcinoma.

The prognostic significance of the detection of HPV (human papilloma virus) DNA in cervical carcinoma was evaluated in 223 cases treated from January 1988 to November 1989. HPV DNA was detected by PCR (polymerase chain reaction) on fresh tumour specimens obtained before therapy was started. HPV DNA of any type was detected in 93.3% of all tumours, HPV16 was the predominant type and was detected in 69% of cases. HPV18 was more frequent in adeno- and adenosquamous carcinoma than in squamous cell carcinoma and occurred more often in poorly differentiated tumours than in more highly differentiated tumours. Patients with HPV negative tumours were on average older than patients with tumours containing HPV. Neither presence of HPV DNA nor HPV type had prognostic significance. In 63 women with early stage tumours submitted to surgery, no difference was found in the frequency of lymph node metastasis, vessel invasion or prognosis related to HPV type. We conclude that neither the presence nor the type of HPV DNA had any prognostic significance in cervical carcinoma.

High incidence of human papillomavirus in 146 cervical carcinomas. A study using three different pairs of consensus primers, and detecting viral genomes with putative deletions.

Polymerase chain reaction (PCR) primer sets and probe-cocktails were used for human papillomavirus (HPV) detection and typing of 146 fresh frozen biopsies of cervical carcinoma. We obtained a high detection rate (96%) by using three sets of consensus primer pairs directed at the L1 and E1 regions of HPV and by probing with a cocktail of random-labelled consensus and type-specific PCR products derived from HPV plasmids. In addition, we performed type-specific PCR amplification with E6-E7 primers. The procedure was designed to detect all HPV-positive cases in a rapid, sensitive and specific way. In addition, by using different regions for amplification, we detected cases with putative genomic deletions in HPV. All the negative PCR and DNA isolation controls were negative. The six negative samples were negative with all probe-cocktails and type-specific primers and three of these negative samples were clear cell carcinomas. The detection rate was similar in squamous carcinomas and in adenocarcinomas and type 16 was most common (65%) in both types of carcinoma. There were no double infections of human papillomavirus 16 and 18.

Human papillomavirus findings in relation to cervical intraepithelial neoplasia grade: a study on 476 Stockholm women, using PCR for detection and typing of HPV.

Among human papillomavirus (HPV) types found in humans, there is a strong association between HPV 16, 18, 31, and 33, and cervical cancer. The relationship between various grades of dysplasia and HPV type is less clear. To elucidate this point, the authors tested 476 cytological and histological samples from cervix with polymerase chain reaction (PCR) for HPV using consensus primer My 09-11 and type-specific primers. All cases were divided into groups on the basis of cytology: "normal cases" (ie, women with other disease than cervical intraepithelial neoplasia [CIN]), and CIN I, II, and III. Out of the "normal cases," in which women with a previous history of condyloma and dysplasia were included, 69% had HPV with type 6 as the most common one. Of all CIN I cases, 71% were HPV positive, and HPV type 6 and 16 were equally common. In CIN II cases, HPV 16 was the most common type, whereas HPV 6 accounted for only 7.5% of cases in this group as single virus type. HPV 16 was also the most common type in the CIN III group, followed by type 33. Double and even multiple infections occurred in all groups.

Presence of human papillomavirus in squamous cell laryngeal carcinomas. A study of thirty-nine cases using polymerase chain reaction and in situ hybridization.

Human papillomavirus (HPV) has a well-established association with benign and malignant cervical tumours as well as with laryngeal papillomas, while its association with laryngeal carcinomas is still controversial. We have examined 39 squamous cell carcinomas of the larynx for HPV using PCR with three consensus primers (Cp, My 09/11 and Gp 5+/6+). Specimens positive for consensus primers were examined with specific PCR primers for HPV 6, 11, 16, 18, 31, 33 and 35 and with in situ hybridization (ISH). Three samples (8%) were found HPV positive for two or three consensus primers. One case was confirmed to be HPV 16 with both PCR and ISH. The findings of both a punctate and diffuse signal with the use of ISH indicated HPV to be in both integrated and episomal form. For the other two HPV positive specimens, the examination indicated a type related to HPV 6/11, but it could not be determined with certainty. This study strongly indicates that HPV 16 is associated with some squamous cell laryngeal carcinomas. The possibility of association with other HPV types or mutants of these needs further investigation.

[Pacemaker failure in elective DC cardioversion of atrial tachycardia]. / Pacemakersvigt ved elektiv DC-konvertering af atrial takykardi.

A 74-year-old woman with a VVI-rate-responsive pacemaker (Pacesetter 2033K; unipolar pacing electrode) was admitted for cardioversion of atrial tachycardia. Antiarrhythmic medication included flecainide 100 mg x 2. Electrical defibrillation was followed by transient, but severe nodal bradycardia and pacemaker malfunction characterized by loss of ventricular capture and sensing. The incident probably represents an example of pacemaker failure due to an acute increase in the stimulation threshold, most likely caused by current-induced tissue damage at the electrode-endomyocardial interface. Flecainide might have contributed to the increase in stimulation threshold. The clinician should be prepared for the possible consequences of pacemaker failure after external defibrillation.

Hands-free sample preparation platform for nucleic acid analysis.

A Lab-On-Chip system with an instrument is presented which is capable of performing total sample preparation and automated extraction of nucleic acid from human cell samples fixed in a methanol based solution. The target application is extraction of mRNA from cervical liquid based cytology specimens for detection of transformed HPV-infections. The device accepts 3 ml of sample and performs the extraction in a disposable polymer chip of credit card size. All necessary reagents for cell lysis, washing, and elution are stored on-chip and the extraction is performed in two filter stages; one for cell pre-concentration and the other for nucleic acid capture. Tests performed using cancer cell lines and cervical liquid based cytology specimens confirm the extraction of HPV-mRNA by the system.

Human papillomavirus E6/E7 mRNA expression in women younger than 30 years of age.

OBJECTIVE: The prevalence of human papillomavirus (HPV) is high in women younger than 30 years of age, most infections being transient. It is not clear, however, to what extent the E6/E7 transcripts are being expressed. This may be of prognostic importance. In this study, we have determined the prevalence of HPV DNA and mRNA in 283 women younger than 30 years of age. METHODS: E6/E7 transcripts from HPV types 16, 18, 31, 33 and 45 were detected using PreTect HPV-Proofer, while the presence of HPV DNA was detected using Gp5+/6+ consensus PCR and type-specific PCR. RESULTS: A total of 92 women (32.5%) were positive by consensus PCR, 59 (20.8%) were positive by type-specific PCR, while 41 (14.5%) were positive by PreTect HPV-Proofer. E6/E7 mRNA expression was detected in 38 (64.4%) of the 59 HPV type-specific DNA positive women. For HPV 16, E6/E7 mRNA expression was observed in 8 (32%) of the 25 DNA positive women. No high-grade lesions were observed in the concomitant cytology. CONCLUSIONS: Among young women having a normal Pap smear, a high HPV prevalence was found. Hence, use of consensus PCR will most probably give a low prognostic value for identifying subsequent severe dysplasia. The five HPV types 16, 18, 31, 33 and 45 accounted for the majority of infections with two out of three having a detectable E6/E7 mRNA expression. Yet, repeated type-specific testing for HPV mRNA may identify young women with a persistent transforming infection being at increased risk for severe dysplasia.

In situ hybridization with nonisotopic probes using different detection systems.

In order to evaluate a sensitive nonisotopic in situ hybridization method for routine work in pathology laboratories, we compared seven different detection systems, using digoxigenin- and biotin-labeled probes. The sensitivity of these methods was tested on four cases of cervical condyloma all known to be positive for HPV 6. Four of these methods gave satisfactory results without any background staining. The single biotin method and the single digoxigenin method were equally sensitive, while the two triple biotin methods, using mouse anti-biotin/anti-mouse IgG/alkaline phosphatase mouse anti-alkaline phosphatase or mouse anti-biotin/alkaline phosphatase anti-mouse IgG/alkaline phosphatase mouse anti-alkaline phosphatase as the detection systems, tremendously improved the sensitivity. The enhanced sensitivity of the nonisotopic in situ hybridization method make it useful in investigation of pathologic tissues.

Dynamics of intrarenal pressures and glomerular filtration rate after acetazolamide.

The dynamics of intrarenal pressures, early distal tubular fluid conductivity (EDC), and renal flood flow (RBF) were studied in rats given acetazolamide (ACZ), an inhibitor of proximal reabsorption. Glomerular filtration rate (GFR) and end-proximal flow were estimated by clearances of 51Cr-EDTA and lithium. Proximal tubular pressure (Pprox) increased initially by 1.7 +/- 0.1 mmHg after ACZ, causing a decrease in the hydrostatic pressure difference across the glomerular membrane (delta P). EDC increased, and then RBF, glomerular capillary pressure (Pgc), Pprox, and star vessel pressures (Psv) dropped as a result of afferent vasoconstriction. Pprox decreased less than Pgc, resulting in a further decrease in delta P, which after 25-30 s reached a constant level 3-4 mmHg below control. After a transient increase the pressures declined to a new steady state, in which Pprox was equal to control, Pgc was decreased, and distal tubular pressure, end-proximal flow, and EDC were increased. GFR was depressed by 29%. The results indicate that the tubuloglomerular feedback mechanism controls Pgc and Pprox by afferent vasoconstriction, as well as efferent vasodilation. The data also indicate that proximal reabsorption rate is important in determining the changes in delta P by its effect on Pprox at least in the early transient phase.

Prevalence and physical state of human papillomavirus DNA in anal carcinomas.

Epidemiologic studies suggest that human papillomavirus (HPV) plays an important role in the development of squamous cell carcinoma of the anal canal. However, in previous studies the prevalence of HPV found in tumor varied from 0 to 85%. This study was performed to investigate the prevalence and physical state of HPV DNA in anal carcinomas. HPV infection was examined using in situ hybridization and polymerase chain reaction. HPV DNA was detected in 80/99 (81%) of the tumors, including 66/74 (89%) from females and 14/25 (56%) from males. HPV16 was demonstrated in 84% of the females and 52% of the males. HPV18, HPV31, and HPV33 could be demonstrated in 5% or less of the cases. In situ hybridization technique showed that 100% of the virus-infected cases had punctate signal and 67% had in addition a diffuse signal, indicating integrated and episomal virus DNA, respectively. The integrated and episomal HPV DNA coexisted in the majority of the tumor cell nuclei. This study indicates that infection and integration of HPV DNA may play an important role in the pathogenesis of anal canal carcinomas.