Reconsideration of the diagnostic criteria required for PCV2 reproductive disease.

Porcine circovirus type 2 (PCV2) causes a variety of clinical conditions including PCV2-associated reproductive disease (PCV2-RD) characterized by late term abortions and mummifications. The generally accepted diagnostic triad includes the presence of reproductive disorders, the histopathological finding of myocarditis, and detection of moderate to high viral loads within the heart tissue. A new threshold of 109 PCV2 genome equivalents (GE)/g heart tissue is suggested to fulfil the third criterion using the diagnostic settings of quantitative real time PCR and in situ hybridization of 30 fetal heart tissues. The need to identify histopathological lesions in fetal heart tissue appears to be invalid or overestimated in confirming a diagnosis of PCV2-RD, at least at the individual fetus level. The highest viral loads (1012 GE/g tissue) were detected in autolyzed and mummified piglets and were identified as PCV2d, although concurrent detection of PCV2d + a and PCV2d + b also occurred.

Low prevalence of porcine circovirus type 2 infections in farrowing sows and corresponding pre-suckling piglets in southern German pig farms.

Porcine circovirus type 2 (PCV2) is the assumed causative agent of a number of different diseases summarized as porcine circovirus diseases (PCVD). The virus is shed via different se- and excretions of PCV2 infected pigs. Transmission of the virus occurs horizontally and vertically either by oronasal or diaplacental infection. Recent research emphasizes the importance of diaplacental PCV2 infection or the infection in early stages of the piglet's life attributable to excretion of PCV2 by the dams within the suckling period. To estimate the prevalence of intrauterine PCV2 infections under field conditions in Bavaria the PCV2 status of farrowing sows (n=198) and corresponding pre-suckling piglets (n=590) of 20 piglet producing farms was examined. PCV2 viral load and anti-PCV2 antibodies in the serum of the sows and piglets were examined at time of farrowing or before colostrum intake, respectively. PCV2 excretion of the sows via saliva, feces and urine was examined additionally. PCV2 specific antibodies in the serum of the sows were detectable on 11 farms with a mean in herd seroprevalence of 35.5% in these farms. Only 0.65% of all samples collected from 198 sows were positive for PCV2 DNA (serum: 1%; feces: 0.5%; saliva: 0.5%; urine: 0.6%). PCV2 DNA was detectable in sample material from seronegative sows as well as from seropositive sows. In none of the pre-suckling serum samples of the piglets IgG antibodies against PCV2 or PCV2 DNA were present. No correlation between the antibody- and viremia status of the sows and the PCV2 excretion was detectable. In contrast to reports about a high prevalence of viremic pre-suckling piglets in the suckling period in North America, the results of the present study reveal that diaplacental infection with PCV2 is comparatively rare in Southern Germany and infection of piglets within the suckling period seems to be more likely.

Detection of a new cluster of porcine circovirus type 2b strains in domestic pigs in Germany.

PCV2 can be divided into three different genotypes: PCV2a, PCV2b and PCV2c. Since 2004/2005 PCV2b has become the predominant genotype in the domestic pig population worldwide. In the years 2010 and 2012 PCV2b mutant strains (mPCV2), classified as PCV2b-1C strains, were detected in porcine circovirus diseases (PCVD) affected pigs in China and the United States, respectively. Within one year (April 2013-April 2014) newly emerging mPCV2 strains were isolated in seven German pig farms routinely vaccinating against PCV2. Histopathological, clinical and molecular biological findings including in-situ hybridization (ISH) and real-time PCR indicate PCVD in the affected animals. Characterized isolates from five farms were closely related to the PCV2b-1C reference strain BDH (GenBank no. HM038017), whereas strains from two other farms were only 99.1% and 99.0% identical (based on the nucleotide sequence of the complete genome) to mPCV2 strain BDH, respectively.

[Unwanted side effects of antibacterials--a diagnostic challenge]. / Unerwünschte Arzneimittelwirkungen von Antibiotika--eine differenzialdiagnostische Herausforderung.

We present three cases of rare side effects which appeared to be attributable to antibacterial drug treatment. A 57-year-old female patient was admitted to hospital due to increasing dyspnea. Computed tomography revealed interstitial lung fibrosis which was attributed to the toxic effects of nitrofurantoin (50 mg/d) that the patient used for approximately one year for recurrent urinary tract infection. She died two weeks after hospital admission due to acute respiratory failure. A 20-year-old male patient presented with most intense headache and psychomotor deceleration. Pseudotumor cerebri, which was suspected to be the underlying cause, is described as a rare side effect of minocyclin which the patient has taken for acne pustulosa (100 mg single dose). After dechallenge of minocyclin, neurological symptoms quickly subsided. A 82-year-old female patient used moxifloxacin (400 mg/d) for febrile bronchopulmonary infection for one week. During this therapy, confusion and severe dementia presented and remained for more than two months after discontinuation. The demential syndrome appears to be possibly related to the fluoroquinolone use. In summary, adverse drug effects not pertaining to the primary physician's field are especially difficult to identify. Most importantly, rare side effects must be borne in mind by the prescribing physician.

Propafenone for the prevention of atrial tachyarrhythmias after cardiac surgery: a randomized, double-blind placebo-controlled trial.

We studied the efficacy of propafenone in preventing atrial tachyarrhythmias after cardiac surgery, and the possible relationships between CYP2D6 polymorphism and the efficacy, pharmacokinetics, and tolerability of propafenone. One hundred and sixty patients were randomized (double blind) to receive propafenone (n= 78) or placebo (n= 82) for 1 week after cardiac surgery. The patients who were assigned to the propafenone group received 1 mg/kg infused in 1 h, followed by a continuous infusion at a rate of 4 mg/kg/24 h until the following morning, and subsequently 450 mg/day orally until the sixth postoperative day. Thirty-seven patients completed the trial in the propafenone group and 45 in the placebo group. The frequency of occurrence of atrial tachyarrhythmia was lower in the propafenone group than in the placebo group (29.7% vs. 53.3%, P< 0.05; relative risk, 0.56). Plasma propafenone concentrations were markedly influenced by CYP2D6 genotype-derived phenotype.

Multiple drug prescribing by general practitioners in a German region: Implications for drug interactions and patient safety.

OBJECTIVE: An increased number of drugs used by patients enhances the risk of potentially hazardous drug interactions. So far, no representative data are available about how common this problem is in German general practices. METHODS: We performed a retrospective analysis using a prescription database for a German region. The 50 general practitioners (out of 1,457) who wrote the most prescriptions during January to March 2003 were included. Data on 4,153 patients who were prescribed at least 10 drugs were analyzed for 92 predefined Drug Combinations Prone to Interact (DCPI) to a clinically relevant extent and possible contraindications. RESULTS: From 92 DCPIs, 71 occurred in the analyzed population between 1 and 275 times. The total number of DCPI cases was 1,295, which included 10% (n = 129) of contraindicated combinations. Among 4,153 analyzed patients, 822 patients (19.8%) were affected by at least 1 DCPI. In 268 patients (6.5%), multiple DCPIs occurred. The most frequently found drug pairs were digitalis/diuretics, digitalis/calcium channel blockers, and theophylline/quinolones. Among contraindicated combinations, tricyclic antidepressants, St. John's wort and antiarrhythmic drugs were most frequently involved. In about 1/3 of patients treated for chronic heart failure, pharmacotherapy appeared not to be guideline-adherent. CONCLUSION: Drug interactions, especially in polypharmacotherapy, represent a potential hazard which must be taken into account by the prescribing physician. Our study is the first to use a prescription database for the evaluation of drug prescriptions within a German region.

Effect of acute hyperhomocysteinemia on methylation potential of erythrocytes and on DNA methylation of lymphocytes in healthy male volunteers.

Homocysteine is a precursor of S-adenosylmethionine (AdoMet) and a metabolite of S-adenosylhomocysteine (AdoHcy). The ratio of AdoMet to AdoHcy, defined as the methylation potential (MP), indicates the flow of methyl groups within the cells. Chronic elevations of total homocysteine (tHcy) in plasma correlate with increased AdoHcy concentrations, decreased MP, and impaired DNA methylation. However, the influence of acute hyperhomocysteinemia on MP is unknown. We induced acute hyperhomocysteinemia in 14 healthy volunteers by oral administration of l-homocysteine (65.1 micromol/kg body wt) in an open, randomized, placebo-controlled two-period crossover study. The kinetics of tHcy in blood and urine, MP in blood, and global DNA methylation in lymphocytes were studied systematically during 48 h. Plasma tHcy concentrations reached a peak at 34 +/- 11 min after an oral load with l-homocysteine and decreased with a half-life of 257 +/- 41 min (means +/- SD). Only 2.3% of the homocysteine dose were recovered in urine. AdoHcy concentrations and MP in whole blood and erythrocytes were not affected by the oral homocysteine load. Furthermore, global DNA methylation in lymphocytes did not change under these conditions. We found no difference between the genotypes of 5,10-methylenetetrahydrofolate reductase in response to the homocysteine load. However, AdoMet content in erythrocytes was significantly higher in the C677T carriers (CT; n = 7) compared with the CC genotype (n = 7). Although chronic elevation of tHcy has been shown to affect MP and DNA methylation, acute elevation of plasma tHcy above 20 micromol/l for 8 h is not sufficient to change MP and to induce DNA hypomethylation in lymphocytes.

Effect of urea and osmotic cell shrinkage on Ca2+ entry and contraction of vascular smooth muscle cells.

The present study was performed to elucidate the effects of urea on vascular smooth muscle cells (SMC). Addition of urea (20, 50, 100 mM) to physiological salt solution blunted the vasoconstrictory effect of phenylephrine (by 17, 25 and 30%, respectively) and of an increased extracellular K+ concentration (by 7, 14 and 19%, respectively) without affecting the basal tone of rabbit arterial rings. According to Fura-2 fluorescence in cultured SMC (A7r5), urea had no effect on basal intracellular calcium activity ([Ca2+]i), but significantly blunted the increase of [Ca2+]i following an increase of extracellular K+. Whole-cell patch-clamp studies revealed that the Ca2+ current through voltage-sensitive Ca2+ channels is significantly inhibited in the presence of urea. As evident from calcein fluorescence, addition of urea leads to sustained cell shrinkage. The effects of urea on vascular tone, [Ca2+]i activity, voltage-gated Ca2+ channels and cell volume are mimicked by addition of raffinose or NaCl. However, the cell shrinkage induced by urea is sustained, whereas the addition of equiosmolar NaCl is only transient and followed by a regulatory cell volume increase. Moreover, hypertonic NaCl increases, whereas urea decreases, the transcription of cell-volume-regulated kinase hsgk. In conclusion, urea leads to sustained shrinkage of vascular smooth muscle cells, which is followed by inhibition of voltage-gated Ca2+ channels, a decrease of [Ca2+]i and thus blunts the vasoconstrictory action of phenylephrine and increased extracellular K+ concentration.