PCR amplification of DNA from malaria parasites on fixed and stained thick and thin blood films

Under some circumstances, polymerase chain reaction (PCR) amplification of deoxyribonucleic acid (DNA) from Plasmodium may become necessary from infections for which only blood slides are available. Established methods used for DNA preparation do not work in that case. We have developed a reliable and controlled method for DNA preparation from malaria parasites on fixed and stained blood films. 162 slides from 2 different locations, some stored for at least one year, have been analysed by PCR amplification of the polymorphic loci for MSA1 and MSA2. In 92% of microscopically positive slides, a PCR product could be detected using material derived from thick blood films. When thin blood films with scanty parasitaemia were used, a PCR product could be obtained with only 71% of samples. In all unsuccessful cases, DNA preparation was the limiting factor, which was controlled for by amplification of a control human templat

Neue Kleeblattkriterien fürs Fleckvieh

Im Stierenkatalog von Swissgenetics werden speziell für Biobetriebe geeignete Stiere mit dem Biokleeblatt ausgezeichnet. Eine Gruppe Bioviehzüchter der Rassen Simmentaler, Swiss Fleckvieh und Red Holstein hat mit Swissgenetics die Kriterien für das Biokleeblatt überarbeitet

Rapid Dynamics of Polyomavirus Type BK in Renal Transplant Recipients

BackgroundPolyomavirus type BK-associated nephropathy (PVAN) is an emerging cause of early renal transplant failure. No specific antiviral treatment has been established. Current interventions rely on improving immune functions by reducing immunosuppression. In patients with PVAN, a high BK virus (BKV) load is detectable in plasma. However, the relationship between BKV replication and disease is not well understood MethodsIn a retrospective analysis of BKV plasma load in renal transplant recipients undergoing allograft nephrectomy (n=3) or changes in immunosuppressive regimen (n=12), we calculated viral clearance rates and generation times and estimated the loss of BKV-infected renal cells ResultsAfter nephrectomy, BKV clearance was fast (viral half-life [t 1/2], 1-2 h) or moderately fast (t 1/2, 20-38 h), depending on the sampling density, but it was independent of continued immunosuppressive regimens. After changing immunosuppressive regimens, BKV was cleared with a t 1/2 of 6 h-17 days. Using the basic reproductive ratio, the efficacies of intervention ranged from 7% to 83% (mean, 28%; median, 22%) ConclusionThe results emphasize that high-level BKV replication is a major pathogenetic factor that may have implications for genome rearrangements, immune evasion, and antiviral resistanc

Randomized, open-label, phase 1/2a study to determine the maximum tolerated dose of intraventricular sustained release nimodipine for subarachnoid hemorrhage (NEWTON [Nimodipine Microparticles to Enhance Recovery While Reducing Toxicity After Subarachnoid Hemorrhage])

BACKGROUND AND PURPOSE—: We conducted a randomized, open-label, phase 1/2a, dose-escalation study of intraventricular sustained-release nimodipine (EG-1962) to determine safety, tolerability, pharmacokinetics, and clinical effects in aneurysmal subarachnoid hemorrhage. METHODS—: Subjects with aneurysmal subarachnoid hemorrhage repaired by clipping or coiling were randomized to EG-1962 or enteral nimodipine. Subjects were World Federation of Neurological Surgeons grade 2 to 4 and had an external ventricular drain. Cohorts of 12 subjects received 100 to 1200 mg EG-1962 (9 per cohort) or enteral nimodipine (3 per cohort). The primary objective was to determine the maximum tolerated dose. RESULTS—: Fifty-four subjects in North America were randomized to EG-1962, and 18 subjects were randomized to enteral nimodipine. The maximum tolerated dose was 800 mg. One serious adverse event related to EG-1962 (400 mg) and 2 EG-1962 dose-limiting toxicities were without clinical sequelae. There was no EG-1962-related hypotension compared with 17% (3/18) with enteral nimodipine. Favorable outcome at 90 days on the extended Glasgow outcome scale occurred in 27/45 (60%, 95% confidence interval 46%–74%) EG-1962 subjects (5/9 with 100, 6/9 with 200, 7/9 with 400, 4/9 with 600, and 5/9 with 800 mg) and 5/18 (28%, 95% confidence interval 7%–48%, relative risk reduction of unfavorable outcome; 1.45, 95% confidence interval 1.04–2.03; P=0.027) enteral nimodipine subjects. EG-1962 reduced delayed cerebral ischemia (14/45 [31%] EG-1962 versus 11/18 [61%] enteral nimodipine) and rescue therapy (11/45 [24%] versus 10/18 [56%]). CONCLUSIONS—: EG-1962 was safe and tolerable to 800 mg, and in this, aneurysmal subarachnoid hemorrhage population was associated with reduced delayed cerebral ischemia and rescue therapy. Overall, the rate of favorable clinical outcome was greater in the EG-1962-treated group. CLINICAL TRIAL REGISTRATION—: URL: http://www.clinicaltrials.gov. Unique identifier: NCT01893190

Acute Renal Failure on Immune Reconstitution in an HIV-Positive Patient with Miliary Tuberculosis

Immune reconstitution syndrome following HAART in human immunodeficiency virus (HIV)-infected patients is characterized by inflammatory worsening of organ functions despite improvement in HIV surrogate markers of HIV infection. We describe a patient with miliary tuberculosis and urinary shedding of acid fast bacilli who developed acute renal failure 8 weeks after initiation of antituberculosis therapy and 6 weeks after initiation of HAART. The diagnostic workup and further course of disease implicated immune reconstitution syndrome as the cause of acute renal failur