GIRK4 confers appropriate processing and cell surface localization to G-protein-gated potassium channels

GIRK1 and GIRK4 subunits combine to form the heterotetrameric acetylcholine-activated potassium current (IKACh) channel in pacemaker cells of the heart. The channel is activated by direct binding of G-protein Gbetagamma subunits. The GIRK1 subunit is atypical in the GIRK family in having a unique ( approximately 125-amino acid) domain in its distal C terminus. GIRK1 cannot form functional channels by itself but must combine with another GIRK family member (GIRK2, GIRK3, or GIRK4), which are themselves capable of forming functional homotetramers. Here we show, using an extracellularly Flag-tagged GIRK1 subunit, that GIRK1 requires association with GIRK4 for cell surface localization. Furthermore, GIRK1 homomultimers reside in core-glycosylated and nonglycosylated states. Coexpression of GIRK4 caused the appearance of the mature glycosylated form of GIRK1. [35S]Methionine pulse-labeling experiments demonstrated that GIRK4 associates with GIRK1 either during or shortly after subunit synthesis. Mutant and chimeric channel subunits were utilized to identify domains responsible for GIRK1 localization. Truncation of the unique C-terminal domain of Delta374-501 resulted in an intracellular GIRK1 subunit that produced normal IKACh-like channels when coexpressed with GIRK4. Chimeras containing the C-terminal domain of GIRK1 from amino acid 194 to 501 were intracellularly localized, whereas chimeras containing the C terminus of GIRK4 localized to the cell surface. Deletion analysis of the GIRK4 C terminus identified a 25-amino acid region required for cell surface targeting of GIRK1/GIRK4 heterotetramers and a 25-amino acid region required for cell surface localization of GIRK4 homotetramers. GIRK1 appeared intracellular in atrial myocytes isolated from GIRK4 knockout mice and was not maturely glycosylated, supporting an essential role for GIRK4 in the processing and cell surface localization of IKACh in vivo.     

Evaluation of a multicomponent, behaviorally oriented, problem-based "summer school" program for adolescents with diabetes

A 2-week summer school program, combining problem-based learning with behavior therapy, was developed to help adolescents with insulin-dependent diabetes improve their ability to cope with obstacles to dietary management. Ten students participated in a first session, and 9 participated in a second session, serving as a waiting list control group. Outcomes were evaluated pre- and postsession and at a 4-month follow-up using 3-day food diaries, blood glucose data, and paper-and-pencil tests of diabetes-related knowledge, self-efficacy, coping strategies, and general problem solving. Improvements were observed in self-efficacy, problem-solving skills, and self-reported coping strategies. No significant changes were observed in daily intake of fat, cholesterol, calories, mean blood glucose levels or blood glucose variability, and diabetes knowledge. Comparisons between the first group and the waiting list control group do not allow the significant pre-post changes to be clearly attributed to the summer school program.     

Geographic variation in the calls of wild chimpanzees: a reassessment

Male chimpanzees produce a species-typical call, the pant hoot, to communicate to conspecifics over long-distances. Calls given by males from the well-known Gombe and Mahale populations typically consist of four different phases: an introduction, build-up, climax, and let-down. Recent observations suggest that chimpanzees living in the Kibale National Park, Uganda, consistently give calls that lack a build-up and are thus qualitatively distinguishable acoustically from those made by other East African conspecifics. We analyzed additional recordings from Mahale and Kibale to re-examine geographic variation in chimpanzee calls. Results indicate that males from both sites produce pant hoots containing all four parts of the call. Calls made by chimpanzees from the two populations, however, differ in quantitative acoustic measures. Specifically, males at Kibale initiate their calls with significantly longer elements and build-up over briefer periods at slower rates than individuals from Mahale. Kibale males also deliver acoustically less variable calls than chimpanzees at Mahale. Although climax elements do not differ between populations in any single acoustic feature, discriminant function analysis reveals that acoustic variables can be used in combination to assign calls to the correct population at rates higher than that expected by chance. Ecological factors related to differences in habitat acoustics, the sound environment of the local biota, and body size are likely to account for these observed macrogeographic variations in chimpanzee calls.     

Adipose tissue development during early postnatal life in ewe-reared lambs

This study examines the precise time course that brown adipose tissue (BAT) takes to adopt the characteristics of white adipose tissue in postnatal lambs. Perirenal adipose tissue was sampled from ewe-reared lambs within 1 h of birth and at 1, 2, 4, 7, 14, 21 and 30 days of age and analysed for the amount of mRNA for uncoupling protein (UCP), the amount and activity of UCP, and protein, mitochondrial protein and lipid content. This was combined with measurements of colonic temperature and jugular venous plasma concentrations of thyroid hormones and insulin-like growth factor-1 (IGF-1). Over the first 4-7 days of age, large quantities of UCP mRNA were associated with a peak in plasma triiodothyronine concentration at 2 days of age followed by a maximal amount and activity of UCP at 4 days and a basal colonic temperature of 39.3 degrees C. Between 7 and 30 days there was a large increase in lipid deposition as the amount and activity of UCP and the amount of UCP mRNA declined to basal values and colonic temperature was maintained at 40 degrees C. A significant positive relationship between perirenal adipose tissue lipid content and plasma IGF-1 concentration was observed throughout the study period. It is concluded that ovine adipose tissue maturation occurs in two distinct phases over the first month of life. The precise time scale of this process could be regulated in part by the lamb's body temperature which determines whether adipose tissue is required for heat production (i.e. BAT) or as an endogenous energy source (i.e. white adipose tissue).     

A synovial sarcoma with a complex t(X;18;5;4) and a break in the ornithine aminotransferase (OAT)L1 cluster on Xp11.2

In this case-control study, we investigated the role of Cryptosporidium in gastroenteritis in children < 6 years old. Six hundred fresh stool specimens were examined for various pathogenic parasites, bacteria, and rotaviruses. Wet-mount preparations, formaline-ether concentrations, and Sheather's floatation techniques were used to recover the parasite oocysts. Permanent stained slides using acid-fast stain and trichrome stains were prepared. Of 300 children with gastroenteritis symptoms, 20 (6.7%) had Cryptosporidium oocysts; seven of the 20 had concomitant infections so they were excluded from the counts. This infection rate is significantly different (Z = 2; p < 0.05) from that found in the control group (1.7%) of children who reported no symptoms. The most frequent symptoms reported beside diarrhea were abdominal pain, cramps, anorexia, nausea, vomiting, and fatigue. Contaminated drinking water is suspected to be the source of infection; other possible factors are discussed.     

Clinical evaluation of low back pain

To properly diagnose and treat low back pain, a thorough history and physical examination are the cornerstones. The most important diagnoses for the physician to be aware of are cauda equina syndrome, back strain, herniated disc, stenosis, and spondylolisthesis.     

Incremental value of myocardial perfusion scintigraphy in prognosis and outcomes of patients with coronary artery disease

The concept of incremental value in prognosis and outcome of patients with coronary artery disease is important to the field of noninvasive imaging. Because these tests are expensive, they should be held to the standard of demonstrating a statistical improvement over the information provided by clinical assessment and treadmill testing. Responding to the demand for cost-effective applications of myocardial perfusion scintigraphy, a large amount of research has recently been devoted to defining specific patient subsets in which incremental value exists for scintigraphy. Subsets thus far demonstrated to benefit incrementally include those men and women referred for possible coronary artery disease, with known coronary artery disease, and after percutaneous transluminal coronary angioplasty, coronary artery bypass grafting, unstable angina, or recent infarction. Incremental cost savings also apply to these subsets except for patients with normal ECGs at rest and less than 15% likelihood for significant coronary artery disease.