Evidence for two independent associations with type 1 diabetes at the 12q13 locus.

Genome-wide association studies have identified associations between type 1 diabetes and single-nucleotide polymorphisms (SNPs) at chromosome 12q13, surrounding the gene ERBB3. Our objective was to fine map this region to further localize causative variants. Re-sequencing identified more than 100 putative SNPs in an 80-kb region at 12q13. By genotyping 42 SNPs, spanning ∼214 kb, in 382 affected sibling pair type 1 diabetes families, we were able to genotype or tag 67 common SNPs (MAF≥0.05) identified from HapMap CEU data and CEU data from the 1000 Genomes Project, plus additional rare coding variants identified from our re-sequencing efforts. In all, 15 SNPs provided nominal evidence for association (P≤0.05), with type 1 diabetes. The most significant associations were observed with rs2271189 (P=4.22 × 10(-5)), located in exon 27 of the ERBB3 gene, and an intergenic SNP rs11171747 (P=1.70 × 10(-4)). Follow-up genotyping of these SNPs in 2740 multiplex type 1 diabetes families validated these findings. After analyzing variants spanning more than 200 kb, we have replicated associations from previous GWAS and provide evidence for novel associations with type 1 diabetes. The associations across this region could be entirely accounted for by two common SNPs, rs2271189 and rs11171747.

A phase I study evaluating the role of the anti-epidermal growth factor receptor (EGFR) antibody cetuximab as a radiosensitizer with chemoradiation for locally advanced pancreatic cancer.

PURPOSE: (1) To determine the safety of the epidermal growth factor receptor (EGFR) antibody cetuximab with concurrent gemcitabine and abdominal radiation in the treatment of patients with locally advanced adenocarcinoma of the pancreas. (2) To evaluate the feasibility of pancreatic cancer cell epithelial-mesenchymal transition (EMT) molecular profiling as a potential predictor of response to anti-EGFR treatment. METHODS: Patients with non-metastatic, locally advanced pancreatic cancer were treated in this dose escalation study with gemcitabine (0-300 mg/m(2)/week) given concurrently with cetuximab (400 mg/m(2) loading dose, 250 mg/m(2) weekly maintenance dose) and abdominal irradiation (50.4 Gy). Expression of E-cadherin and vimentin was assessed by immunohistochemistry in diagnostic endoscopic ultrasound fine-needle aspiration (EUS-FNA) specimens. RESULTS: Sixteen patients were enrolled in 4 treatment cohorts with escalating doses of gemcitabine. Incidence of grade 1-2 adverse events was 96%, and incidence of 3-4 adverse events was 9%. There were no treatment-related mortalities. Two patients who exhibited favorable treatment response underwent surgical exploration and were intraoperatively confirmed to have unresectable tumors. Median overall survival was 10.5 months. Pancreatic cancer cell expression of E-cadherin and vimentin was successfully determined in EUS-FNA specimens from 4 patients. CONCLUSIONS: Cetuximab can be safely administered with abdominal radiation and concurrent gemcitabine (up to 300 mg/m(2)/week) in patients with locally advanced adenocarcinoma of the pancreas. This combined therapy modality exhibited limited activity. Diagnostic EUS-FNA specimens could be analyzed for molecular markers of EMT in a minority of patients with pancreatic cancer.

Variants in intron 13 of the ELMO1 gene are associated with diabetic nephropathy in African Americans.

Variants in the engulfment and cell motility 1 (ELMO1) gene are associated with nephropathy due to type 2 diabetes mellitus (T2DM) in a Japanese cohort. We comprehensively evaluated this gene in African American (AA) T2DM patients with end-stage renal disease (ESRD). Three hundred and nine HapMap tagging SNPs and 9 reportedly associated SNPs were genotyped in 577 AA T2DM-ESRD patients and 596 AA non-diabetic controls, plus 43 non-diabetic European American controls and 45 Yoruba Nigerian samples for admixture adjustment. Replication analyses were conducted in 558 AA with T2DM-ESRD and 564 controls without diabetes. Extension analyses included 328 AA with T2DM lacking nephropathy and 326 with non-diabetic ESRD. The original and replication analyses confirmed association with four SNPs in intron 13 (permutation p-values for combined analyses = 0.001-0.003), one in intron 1 (P = 0.004) and one in intron 5 (P = 0.002) with T2DM-associated ESRD. In a subsequent combined analysis of all 1,135 T2DM-ESRD cases and 1,160 controls, an additional 7 intron 13 SNPs produced evidence of association (P = 3.5 x 10(-5)- P = 0.05). No associations were seen with these SNPs in those with T2DM lacking nephropathy or with ESRD due to non-diabetic causes. Variants in intron 13 of the ELMO1 gene appear to confer risk for diabetic nephropathy in AA.

Characterization of RNA interference in an Anopheles gambiae cell line.

Introduction of double stranded RNA into invertebrate cells often results in posttranscriptional silencing of target genes through a mechanism termed RNA interference (RNAi). Double-stranded RNA is cleaved by an RNAse III-like enzyme, termed dicer, to small interfering RNAs (siRNAs). In Drosophila, these siRNAs are incorporated in the RNA induced silencing complex (RISC) and mediate degradation of target mRNA. The RISC complex contains members of Argonaute (Ago) family of proteins. We show here that RNAi in a hemocyte cell line of Anopheles gambiae, the principal malaria vector in Africa, requires expression of dicer-2, Ago2 and Ago3 proteins. Furthermore, we demonstrate that RNAi in the mosquito does not spread outside of the target region, suggesting that RNA dependent RNA polymerase mediated transitive amplification is absent in the mosquito.

Angiotensin I-converting enzyme antisense gene therapy causes permanent antihypertensive effects in the SHR.

The renin-angiotensin system plays a critical role in the control of blood pressure (BP), and its hyperactivity is associated with the development and maintenance of hypertension. Although traditional pharmacological therapies targeted toward the inhibition of the renin-angiotensin system are effective in the control of this disease, they pose significant limitations. We used an antisense gene delivery strategy to circumvent these limitations and established that a single intracardiac administration of angiotensin type 1 receptor antisense (AT(1)R-AS) causes permanent prevention of hypertension in the spontaneously hypertensive rat (SHR), an animal model of primary human hypertension. Our objectives in this study were 2-fold: to determine (1) whether the targeting of angiotensin I-converting enzyme (ACE) mRNA by a similar antisense strategy would prevent the SHR from developing hypertension and (2) whether the antihypertensive phenotype is transmitted to the offspring from the antisense-treated parents. Administration of a retroviral vector containing ACE antisense (LNSV-ACE-AS) caused a modest yet significant attenuation of high BP ( approximately 15+/-2 mm Hg) exclusively in the SHR. This was associated with a complete prevention of cardiac and renovascular pathophysiological alterations that are characteristic of hypertension. Like their parents, the F(1) generation offspring of the LNSV-ACE-AS-treated SHR expressed lower BP, decreased cardiac hypertrophy, and normalization of renal arterial excitation-coupling compared with offspring derived from the LNSV-ACE-tS (truncated sense)-treated SHR. In addition, the endothelial dysfunction commonly observed in the SHR renal arterioles was significantly prevented in both parents and offspring of the LNSV-ACE-AS-treated SHR. Polymerase chain reaction followed by Southern analysis revealed that the ACE-AS was integrated into the SHR genome and transmitted to the offspring. These observations suggest that transmission of ACE-AS by retroviral vector may be responsible for the transference of normotensive phenotypes in the SHR offspring.

Angiotensin I-converting enzyme antisense prevents altered renal vascular reactivity, but not high blood pressure, in spontaneously hypertensive rats.

The renin-angiotensin system plays a critical role in the control of blood pressure, and its hyperactivity is associated with the development of human primary hypertension. Because low-dose angiotensin I-converting enzyme (ACE) inhibitors cause small reductions in blood pressure that are associated with the complete reversal of altered vascular pathophysiology, our objective in this study was to determine whether ACE antisense (ACE-AS) gene delivery prevents alterations in renal vascular physiology in the parents and F(1) offspring of AS-treated spontaneously hypertensive rats (SHR). A single bolus intracardiac injection of ACE-AS (2x10(8) colony-forming units) in SHR neonates caused a modest (18+/-3 mm Hg, n=7 to 9) lowering of blood pressure, which was maintained in the F(1) generation offspring (n=7 to 9). Alterations in renal vascular reactivity, electrophysiology, and [Ca(2+)](i) homeostasis are underlying mechanisms associated with the development and establishment of hypertension. Renal resistance arterioles from truncated ACE sense-treated SHR showed a significantly enhanced contractile response to KCl and phenylephrine (n=24 rings from 6 animals, P<0.01) and significantly attenuated acetylcholine-induced relaxations (n=24 rings from 6 animals, P<0.01) compared with arterioles from ACE-AS-treated SHR. In addition, compared with cells dissociated from arterioles of ACE-AS-treated SHR, cells from truncated ACE sense-treated animal vessels had a resting membrane potential that was 22+/-4 mV more depolarized (n=38, P<0.01), an enhanced L-type Ca(2+) current density (2.2+/-0.3 versus 1.2+/-0.2 pA/pF, n=23, P<0.01), a decreased Kv current density (16.2+/-1.3 versus 5.4+/-2.2 pA/pF, n=34, P<0.01), and increased Ang II-dependent changes in [Ca(2+)](i) (n=142, P<0.01). Similar effects of ACE-AS treatment were observed in the F(1) offspring. These results demonstrate that ACE-AS permanently prevents alterations in renal vascular pathophysiology in spite of the modest effect that ACE-AS had on high blood pressure in SHR.

The effect of nicotine on growth of Streptococcus mutans.

There is little information available concerning the effects of nicotine on oral bacteria; and in particular, Streptococcus mutans, an important microbe in the etiology of dental caries. To test the effects of nicotine, Streptococcus mutans were incubated in either 0 or 10(-1)-10(-7) M concentrations of nicotine and then plated onto mitis-salivarius-bacitracin agar. Colonies were counted and treatment groups compared. Both 10(-1) and 10(-2) M nicotine caused total inhibition of bacterial growth, while 10(-3) and 10(-4) M produced significantly more colonies than control. Also, 10(-6) and 10(-7) M nicotine produced a significant reduction in the mean number of colonies. These results suggest a biphasic, dosage-dependent effect of nicotine on the growth of Streptococcus mutans. Since 10(-3) M nicotine has been reported within the saliva of smokeless tobacco users, use of these products could stimulate growth of Streptococcus mutans and possibly place the user at increased risk for dental caries.

Effect of semi-rigid lumbosacral orthosis use on oxygen consumption during repetitive stoop and squat lifting.

The use of back belts in industry has increased despite the lack of scientific evidence supporting their efficacy. The purpose of this study was to investigate the effect of a semi-rigid lumbosacral orthosis (SRLSO) on oxygen consumption during 6-min submaximal repetitive lifting bouts of 10 kg at a lifting frequency of 20 repetitions min-1. Fifteen healthy subjects (13 men, two women) participated in this study. Each subject performed squat and stoop lifting with and without an SRLSO for a total of four lifting bouts. Lifting bouts were performed in random order. Oxygen consumption during the final minute of each lifting bout was used for analysis. A two-way analysis of variance with repeated measures was used to analyse the effects of lift and belt conditions. The stoop and squat methods were significantly different, with the squat lift requiring 23% more oxygen on average than the stoop lift for equal bouts of work. No significant difference was found between the belt and no belt condition within the same lifting technique and no interaction was present. These data suggest that an SRLSO does not passively assist the paravertebral muscles (PVM) in stabilizing the spine during submaximal lifting bouts.

Distance learning: an innovative approach to nursing education.

This article presents four views encompassing one nursing department's experience using distance learning technology. The challenge of presenting a nursing class through distance technology is discussed from the perspective of the telecommunication faculty providing the technological support, the nursing faculty teaching the on-campus course, the off-campus nursing faculty coordinating the course at the outreach site, and the head of the Department of Nursing. Review of course grades demonstrated that off-campus students achieved higher grades than on-campus students. All students evaluated the teacher as being effective; however, off-campus students were not as strong in their opinion.

Electrical muscle stimulation in the treatment of progressive adolescent idiopathic scoliosis: a literature review.

Electrical muscle stimulation is quickly becoming accepted as a reasonable alternative to bracing in the treatment of mild to moderate progressive adolescent idiopathic sciolosis. Stimulation is applied nightly during the eight to ten sleeping hours. With success defined as less than or equal to four degrees of progression, between 60 and 84% of curves treated have been halted. Greater success has been noted in the lumbar and thoracolumbar curves and in curves under 40 degrees. In non-compliant patients, approximately 80% of the curves progressed. Controversy concerning minor refinements in treatment application, such as the details of electrode placement and problems of skin rash and sleep loss, have yet to be satisfactorily resolved.