Insulin capture by a G-quadruplex DNA oligonucleotide containing a two-repeat sequence of the insulin-linked polymorphic region (ILPR) of the human insulin gene promoter region is reported. The immobilized oligonucleotide was demonstrated to capture human insulin from standard solutions and from nuclear extracts of pancreatic cells with high selectivity, using affinity MALDI-mass spectrometry and affinity capillary chromatography. Insulin was preferentially captured by the tworepeat ILPR oligonucleotide over another G-quadruplex forming oligonucleotide, the thrombin binding aptamer, as well as over a single repeat of the ILPR sequence that is not capable of forming the G-quadruplex architecture. Binding was shown to involve the beta chain of insulin, most likely through association with the two parallel loops of the G-quadruplex structure. The discovery raises the possibility that insulin may bind to G-quadruplex DNA formed in the ILPR in vivo and thereby play a role in modulation of insulin gene expression, and provides a basis for design of insulin analogs to probe this hypothesis. The availability of a DNA ligand to human insulin has analytical importance as well, offering an alternative to antibodies for in vitro or in vivo detection and sensing of insulin as well as its isolation and purification from biological samples.

ObjectiveThis report warns that gastrogastric fistulas may follow the division of the stomach in bariatric surgery. Summary Background DataAlthough surgery is the most effective therapy for morbid obesity, the procedures are still undergoing evolution. One of the key elements in bariatric surgery is the partition of the stomach to develop a much smaller reservoir. The partition has been done with single layers of staples with almost universal failure and with double layers of staples with a failure rate of 11.8% when observed for a 12-year follow-up. MethodsThis report details the experience with a series of 100 consecutive patients in whom the partition was created by dividing the stomach. ResultsThe course of six patients was complicated by gastrogastric fistulas. One of the patients had the gastric bypass as the initial bariatric operation; in the other five, the gastric bypasses were carried out to revise failed staple lines. Although one of the patients required drainage for a subphrenic abscess, two had only self-limited signs of infection. In the remaining three patients, there was no evidence of any complication. ConclusionGastrogastric fistulas followed division of the stomach in 6% of our gastric bypass operations.Methods for avoiding this complication include oversewing staple lines, using strong bites of tissue during the anastomosis, avoiding obstruction of the Roux-en-Y jejunal segment, and testing of the integrity of the anastomosis with methylene blue dyes. The ideal method for partition of the stomach remains to be developed.

Canine T cell lymphoma has previously been found to be a poor prognostic indicator compared with its B cell counterpart. The cyclophosphamide, doxorubicin, vincristine, and prednisolone protocol is widely accepted as a first line treatment for canine lymphoma. There have been several studies investigating alternative protocols for T cell lymphoma. This study investigated the use of a modified lomustine, vincristine, procarbazine and prednisolone protocol as a first line treatment in 35 dogs with T Cell lymphoma. Median progression free survival (PFS) time for all 35 dogs was 431 days with a 6-month, 1-year, 2-year, and 3-year PFS of 69%, 54%, 29%, and 12%. Median survival time (MST) was 507 days. Twenty-nine dogs attained a complete response and had a median PFS time of 509 days. Thirty dogs experienced adverse events during the protocol, with 73% of these being grade 1 or 2. This protocol has shown increased median PFS time and MST compared with previous studies and suggests its use as a first line chemotherapy protocol against canine T cell lymphoma.

Capture and detection of Immunoglobulin E (IgE) in simple solution and in human serum using an aptamer-modified probe surface for affinity Matrix-Assisted Laser Desorption-Ionization Mass Spectroscopy (affinity MALDI-MS) detection is reported. Detectable signals were obtained for 1 amol of IgE applied either in a single, 1 μL application of 1 pM IgE or after 10 successive, 1 μL applications of 100 fM IgE. In both cases, the surface was rinsed after each application of IgE to remove sample concomitants including salts and free or non-specifically associated proteins. Detection of native IgE, which is the least abundant of the serum immunoglobulins and occurs at sub-nM levels, in human serum was demonstrated and interference from the high abundance immunoglobulins and albumin was investigated. The aptamermodified surface showed high selectivity towards immunoglobulins in serum, with no significant interference from serum albumin. Addition of IgE to the serum suppressed the signals from the other immunoglobulins, confirming the expected selectivity of the aptamer surface towards IgE. Dilution of the serum increased the selectivity toward IgE; the protein was detected without interference in a 10,000-fold dilution of the serum, which is consistent with detection of IgE at amol (pM) levels in standard solutions.