Poop transplants work as well as antibiotics to treat a common and deadly cause of diarrhea, and perhaps even better, researchers reported Saturday.
Their small and unusual study suggests that doctors should try fecal transplants right off when patients in the hospital get infected with Clostridium difficile, a frequent cause of diarrhea in frail and sick patients...
Heart tissue sections (10 mg) were subjected to Proteinase-K lysis and total DNA was extracted by phenol/chloroform extraction/ethanol precipitation method. Total DNA (100 ng) was treated with RNase A (EN0531, Thermo Scientific, Waltham MA), purified by using DNeasy Mini Spin Columns (Qiagen, Germantown MD), and examined for quality (OD260/OD280 ratio of 1.7–2.0) and quantity ([OD260 –OD320] x 50-?g/ml) by using a DU 800 UV/visible spectrophotometer. To assess the cardiac mtDNA integrity, total DNA was used as template with mtDNA-specific primer sets and PfuUltra II Fusion HS DNA polymerase (Stratagene, La Jolla CA). The long range PCR to amplify 10 kb mtDNA was performed with hot start of 2 [email protected]°C followed by [email protected]°C, [email protected]°C, [email protected]°C for 28 cycles. Densitometry analysis of mtDNA bands was performed as above, and 10 kb mtDNA level was normalized to short-length, 117-bp mtDNA and 96-bp GAPDH nuDNA.
To assess tissue parasite burden, total DNA (100 ng) was used as a template with SYBR Green Supermix (1708882, Bio-Rad) and Tc18SrDNA-specific primers, and real-time qPCR was performed on an iCycler thermal cycler. Data were normalized to GAPDH, and relative parasite burden was calculated as 2-?Ct (?Ct = CtTc18SrDNA—CtGAPDH). The primers are listed in Table B of S1 Table.