NMR studies of caldesmon-calmodulin interactions

The binding of the calcium-regulatory protein calmodulin (CaM) to caldesmon (CaD) contributes to the regulation of smooth muscle contraction. Two regions of caldesmon have been identified as putative calmodulin-binding domains. We have earlier reported on the binding of one of these domains to calmodulin (Zhang & Vogel (1994) Biochemistry 33, 1163-1171). Here we have studied the binding of CaM to synthetic peptides of CaD which contain: (1) both the first and second CaM-binding domains; (2) the second CaM-binding domain; and (3) the sequence between the first and second CaM-binding domains. Two-dimensional transferred nuclear Overhauser enhancement proton NMR measurements as well as circular dichroism studies of a 22-residue peptide NKETAGLKVGVSSRINEWLTK, which contains the second CaM-binding domain, show that only the C-terminal half of the peptide becomes alpha-helical upon binding to CaM. Somewhat surprisingly, the shorter 9-residue peptide SRINEWLTK was sufficient to form a 1:1 complex with CaM; this peptide appears to bind as a 3(10)-helix. Proton-carbon-13 correlation NMR titration studies with specifically labeled [methyl-13C]methionine CaM were used to study the participation of the hydrophobic regions in both domains of the dumbbell shaped CaM in peptide binding. Binding of a 54-residue CaD peptide containing both CaM-binding domains affects all the 8 Met residues in the two hydrophobic domains of CaM (only Met 76 in the linker region of CaM is not involved), while binding of the second CaM-binding domain of CaD influences principally Met 51, 71, and Met 124, 144. Simultaneous binding to CaM of two peptides comprising the first and the second CaM-binding domains also caused changes to all Met residues except Met 76. Taken together, these data demonstrate that both CaM-binding domains of CaD can bind simultaneously to the two hydrophobic regions of CaM.     

Induction of fibrinogen expression in the lung epithelium during Pneumocystis carinii pneumonia

Pneumocystis carinii is an important pulmonary pathogen responsible for morbidity and mortality in patients with AIDS. The acute-phase response (APR), the primary mechanism used by the body to restore homeostasis following infection, is characterized by increased levels of circulating fibrinogen (FBG). Although the liver is the primary site of increased FBG synthesis during the APR, we unexpectedly discovered that FBG is synthesized and secreted by lung alveolar epithelial cells in vitro during an inflammatory stimulus. Therefore, we sought to determine whether lung epithelial cells produce FBG in vivo using animal models of P. carinii pneumonia (PCP). Inflammation was noted by an influx of macrophages to P. carinii-infected alveoli. Northern hybridization revealed that gamma-FBG mRNA increased two- to fivefold in P. carinii-infected lung tissue, while RNA in situ hybridization demonstrated increased levels of gamma-FBG mRNA in the lung epithelium. Immunoelectron microscopy detected lung epithelial cell-specific production of FBG, suggesting induction of a localized inflammatory response resembling the APR. A systemic APR was confirmed by a two- to fivefold upregulation of the levels of hepatic gamma-FBG mRNA in animals with PCP, resulting in a corresponding increase in levels of FBG in plasma. Furthermore, immunoelectron microscopy revealed the presence of FBG at the junction of cell membranes of trophic forms of P. carinii organisms aggregated along the alveolar epithelium. These results implicate FBG in the pathogenesis of PCP in a manner similar to that of the adhesive glycoproteins fibronectin and vitronectin, which are known to participate in intra-alveolar aggregation of organisms and adherence of P. carinii to the lung epithelium.     

Enhancing the performances of recycling folded cascode OpAmp in nanoscale CMOS through voltage supply doubling and design for reliability

Current‐oriented operational amplifier (OpAmp) design has been common for its orderly current‐to‐speed tradeoff. However, for high‐precision or high‐linearity applications, increasing the current does not help much, as the supply voltage (V_DD) and intrinsic gain of the MOSFETs in ultra‐scaled CMOS technologies are very limited. This paper introduces voltage‐oriented circuit techniques to address such limitations. Specifically, a 2xV_DD‐enabled recycling folded cascade (RFC) OpAmp is proposed. It features: (1) current recycling to enhance the effective trans conductance by 4x with no extra power; (2) transistor stacking to boost the output resistance by one to two orders of magnitude; and (3) V_DD elevating to enlarge the linear output swing by 4x. Comparing with its 1xV_DD RFC and FC counterparts, the proposed solution achieves 20‐dB higher DC gain (i.e. 72.8 dB) in open loop and 20‐dB lower IM3 (i.e., –76.5 dB) in closed loop, under the same power budget of 0.6 mW in a 1‐V General Purpose 65‐nm CMOS process. In many applications, these joint improvements in a single stage are already adequate, being more power efficient (i.e. less current paths), stable (i.e. more phase margin), and compact (i.e. no frequency compensation) than multi‐stage OpAmps. Voltage‐conscious biasing and node‐voltage trajectory check ensure the device reliability in both transient and steady states. No specialized high‐voltage device is necessary. Copyright © 2012 John Wiley & Sons, Ltd.     

Dietary patterns during pregnancy and risk of gestational diabetes: a prospective cohort study in Western China

Background

Previous research has demonstrated the association between maternal dietary patterns and gestational diabetes (GDM), but evidence in Asian populations remains limited and inconsistent. This study investigated the association between dietary patterns during early pregnancy and the risk of GDM among pregnant women in Western China.

Methods

A prospective cohort study was conducted among 1337 pregnant women in Western China. Dietary intakes were assessed at 15–20 weeks of gestation using a validated food frequency questionnaire. GDM was diagnosed by oral glucose tolerance tests at 24–28 weeks of gestation. Exploratory factor analysis was performed to derive dietary patterns, and logistic regression models were used to examine the association between dietary patterns and GDM.

Results

A total of 199 women (14.9%) developed GDM. Three dietary patterns were identified, namely, a plant-based pattern, a meat-based pattern and a high protein-low starch pattern. Notwithstanding a lack of association between dietary patterns and GDM risk in the whole cohort, there was a significant reduction in GDM risk among overweight women (BMI ≥24 kg/m²); the odds ratio being 0.29 (95% confidence interval 0.09 to 0.94) when comparing the highest versus the lowest score of the high protein-low starch pattern.

Conclusions

There was no significant association between early pregnancy dietary patterns and GDM risk later in pregnancy for women in Western China, but high protein-low starch diet was associated with lower risk for GDM among women who were overweight at pre-pregnancy.

     

Maximum Penalized Likelihood Kernel Regression for Fast Adaptation

This paper proposes a nonlinear generalization of the popular maximum-likelihood linear regression (MLLR) adaptation algorithm using kernel methods. The proposed method, called maximum penalized likelihood kernel regression adaptation (MPLKR), applies kernel regression with appropriate regularization to determine the affine model transform in a kernel-induced high-dimensional feature space. Although this is not the first attempt of applying kernel methods to conventional linear adaptation algorithms, unlike most of other kernelized adaptation methods such as kernel eigenvoice or kernel eigen-MLLR, MPLKR has the advantage that it is a convex optimization and its solution is always guaranteed to be globally optimal. In fact, the adapted Gaussian means can be obtained analytically by simply solving a system of linear equations. From the Bayesian perspective, MPLKR can also be considered as the kernel version of maximum a posteriori linear regression (MAPLR) adaptation. Supervised and unsupervised speaker adaptation using MPLKR were evaluated on the Resource Management and Wall Street Journal 5K tasks, respectively, achieving a word error rate reduction of 23.6% and 15.5% respectively over the speaker-independently model.     

A novel polymer microneedle fabrication process for active fluidic delivery

In this article, we explore a new fabrication process for a flexible, all polymer, active fluidic delivery system, incorporating a fusion of laser micromachining and microfabrication techniques as well as rapid prototyping technology. Here, we show selective fluidic delivery from isolated microchannels through an electrochemically driven pumping reaction, demonstrate the dispensing of dose volumes up to 5.5 μl, and evaluate the device’s performance in terms of its delivery speed and ejection efficiency. Finally, we move this work toward an implantable microfluidic drug delivery device by investigating the device’s biocompatibility through a statistical approach that overviews the viability of bovine aortic endothelial cells on polyimide and silicon substrates.