Specific and efficient peptide substrates for assaying the proteolytic activity of prostate-specific antigen

Prostate-specific antigen (PSA) is a serine protease secreted by both normal prostate glandular cells and prostate cancer cells. The major proteolytic substrates for PSA are the gel-forming proteins in semen, semenogelin (Sg) I and II. On the basis of the PSA cleavage map for Sg I and II, a series of small peptides (i.e., < or = 7 amino acids) was synthesized and coupled at the COOH terminus to 7-amino-4-methyl coumarin. Using these fluorescently tagged substrates, K(m)s and k(cat)s were determined for PSA hydrolysis, and the substrates were also tested for activity against a panel of purified proteases. Previously, a variety of chymotrypsin substrates have been used to assay the enzymatic activity of PSA. The present studies have identified a peptide sequence with a high degree of specificity for PSA (ie., no detectable hydrolysis by chymotrypsin) and improved K(m)s and k(cat)s over previously used substrates. On the basis of these parameters, the best peptide substrate for PSA has the amino acid sequence HSSKLQ. Using PC-82 human prostate cancer xenografts and human prostate tissues, this PSA substrate was used to document that prostate cancer cells secrete enzymatically active PSA into the extracellular fluid but that once in the blood, PSA is not enzymatically active. On the basis of this information, it should be possible to use the HSSKLQ peptide as a carrier to target peptide-coupled prodrugs for selective activation within sites of PSA-secreting, metastatic prostate cancer cells and not within the blood or other nonprostatic normal tissues.     

Chemical characterization of the predominant proteins secreted by mouse seminal vesicles

Eosinophilic fasciitis is a disorder characterised by induration of the skin due to chronic inflammation and fibrosis of the subcutaneous septa and muscular fascia. It is different from sclerodermia. This is a clinical entity that presents as swelling, tenderness and stiffness of the extremities associated with peripheral eosinophilia. We described a patient with this disorder with 18 month of evolution who had a bilateral carpal tunnel syndrome. Her disease appeared after steroid treatment. A review of medical literature demonstrated that similar clinical pictures are originated by different causes. Some authors propose to encompass this group of disorder under the designation of "fasciitis-panniculitis syndrome" instead of "eosinophilic fasciitis" although the well-known eosinophilic fasciitis is clearly recognized and demonstrated.     

Optical Activation of TrkB (E281A) in Excitatory and Inhibitory Neurons of the Mouse Visual Cortex

The activation of tropomyosin receptor kinase B (TrkB), the receptor of brain-derived neurotrophic factor (BDNF), plays a key role in induced juvenile-like plasticity (iPlasticity), which allows restructuring of neural networks in adulthood. Optically activatable TrkB (optoTrkB) can temporarily and spatially evoke iPlasticity, and recently, optoTrkB (E281A) was developed as a variant that is highly sensitive to light stimulation while having lower basal activity compared to the original optoTrkB. In this study, we validate optoTrkB (E281A) activated in alpha calcium/calmodulin-dependent protein kinase type II positive (CKII⁺) pyramidal neurons or parvalbumin-positive (PV⁺) interneurons in the mouse visual cortex by immunohistochemistry. OptoTrkB (E281A) was activated in PV⁺ interneurons and CKII⁺ pyramidal neurons with blue light (488 nm) through the intact skull and fur, and through a transparent skull, respectively. LED light stimulation significantly increased the intensity of phosphorylated ERK and CREB even through intact skull and fur. These findings indicate that the highly sensitive optoTrkB (E281A) can be used in iPlasticity studies of both inhibitory and excitatory neurons, with flexible stimulation protocols in behavioural studies.     

Microfluidic, label-free enrichment of prostate cancer cells in blood based on acoustophoresis

Circulating tumor cells (CTC) are shed in peripheral blood at advanced metastatic stages of solid cancers. Surface-marker-based detection of CTC predicts recurrence and survival in colorectal, breast, and prostate cancer. However, scarcity and variation in size, morphology, expression profile, and antigen exposure impairs reliable detection and characterization of CTC. We have developed a noncontact, label-free microfluidic acoustophoresis method to separate prostate cancer cells from white blood cells (WBC) through forces generated by ultrasonic resonances in microfluidic channels. Implementation of cell prealignment in a temperature-stabilized (±0.5 °C) acoustophoresis microchannel dramatically enhanced the discriminatory capacity and enabled the separation of 5 μm microspheres from 7 μm microspheres with 99% purity. Next, we determined the feasibility of employing label-free microfluidic acoustophoresis to discriminate and divert tumor cells from WBCs using erythrocyte-lysed blood from healthy volunteers spiked with tumor cells from three prostate cancer cell-lines (DU145, PC3, LNCaP). For cells fixed with paraformaldehyde, cancer cell recovery ranged from 93.6% to 97.9% with purity ranging from 97.4% to 98.4%. There was no detectable loss of cell viability or cell proliferation subsequent to the exposure of viable tumor cells to acoustophoresis. For nonfixed, viable cells, tumor cell recovery ranged from 72.5% to 93.9% with purity ranging from 79.6% to 99.7%. These data contribute proof-in-principle that label-free microfluidic acoustophoresis can be used to enrich both viable and fixed cancer cells from WBCs with very high recovery and purity.     

The nature of the copper sulfide film grown on copper in aqueous sulfide and chloride solutions

In this paper, the properties of copper sulfide films formed both anodically and naturally in deaerated/anoxic aqueous sulfide and chloride solutions were investigated using a series of electrochemical and surface analytical techniques. A combination of cyclic voltammetric, corrosion potential (E_corr), and cathodic stripping voltammetric experiments showed that the sulfide film growth kinetics and film morphologies were controlled by the supply of SH⁻ from the bulk solution to the copper surface. There was no passive barrier layer observed on the copper surface under either electrochemical or corrosion conditions. The film morphology was dependent on the type and concentration of anions (SH⁻, Cl⁻) present in the solution. Scanning electron microscopy on both surfaces and focused ion beam-cut cross-sections showed the growth of a thin, but porous, base layer of chalcocite (Cu₂S) after short immersion periods (up to 2 hr) and the continuous growth of a much thicker crystalline outer deposit over longer immersion periods (≥36 hr), suggesting a solution species transport-based film formation process and the formation of an ineffective thin “barrier-type” layer on copper.     

基于超几何分解的随机运算系统分析方法

基于Bernoulli分布的方差及期望传递方程一直是随机运算系统的数学基础,针对这种传统分析方法在实际应用中的不准确性和片面性,该文提出一种全新的数学方法：超几何分解(hypergeometic decomposition),用来解决在更复杂情况下期望与方差在随机运算系统中的传播规律。基于超几何分解,提出4组更加精确的期望及方差传递方程,在数学上证明了随机运算体系更加广泛的适用性,并且通过随机运算系统在图像处理中的应用,提出了基于方差的系统评价方法,相比于传统按位仿真方法,基于方差的系统分析方法具有耗时短、准确和全面的优点。新的方差传递方程首次将随机信号源的类型引入性能分析,证明了具有特定码流长度的随机序列可以使系统性能达到最优。     

The impact of incorrectly speculated memory operations in a multithreaded architecture

The speculated execution of threads in a multithreaded architecture, plus the branch prediction used in each thread execution unit, allows many instructions to be executed speculatively, that is, before it is known whether they actually needed by the program. In this study, we examine how the load instructions executed on what turn out to be incorrectly executed program paths impact the memory system performance. We find that incorrect speculation (wrong execution) on the instruction and thread-level provides an indirect prefetching effect for the later correct execution paths and threads. By continuing to execute the mispredicted load instructions even after the instruction or thread-level control speculation is known to be incorrect, the cache misses observed on the correctly executed paths can be reduced by 16 to 73 percent, with an average reduction of 45 percent. However, we also find that these extra loads can increase the amount of memory traffic and can pollute the cache. We introduce the small, fully associative wrong execution cache (WEC) to eliminate the potential pollution that can be caused by the execution of the mispredicted load instructions. Our simulation results show that the WEC can improve the performance of a concurrent multithreaded architecture up to 18.5 percent on the benchmark programs tested, with an average improvement of 9.7 percent, due to the reductions in the number of cache misses.     

Performance-based path determination for interprocessorcommunication in distributed computing systems

The different types of messages used by a parallel application program executing in a distributed computing system can each have unique characteristics so that no single communication network can produce the lowest latency for all messages. For instance, short control messages may be sent with the lowest overhead on one type of network, such as Ethernet, while bulk data transfers may be better suited to a different type of network, such as Fibre Channel or HIPPI. This work investigates how to exploit multiple heterogeneous communication networks that interconnect the same set of processing nodes using a set of techniques we call performance-based path determination (PBPD). The performance-based path selection (PBPS) technique selects the best (lowest latency) network among several for each individual message to reduce the communication overhead of parallel programs. The performance-based path aggregation (PBPA) technique, on the other hand, aggregates multiple networks into a single virtual network to increase the available bandwidth. We test the PBPD techniques on a cluster of SGI multiprocessors interconnected with Ethernet, Fibre Channel, and HiPPI networks using a custom communication library built on top of the TCP/IP protocol layers. We find that PBPS can reduce communication overhead in applications compared to using either network alone, while aggregating networks into a single virtual network can reduce communication latency for bandwidth-limited applications. The performance of the PBPD techniques depends on the mix of message sizes in the application program and the relative overheads of the networks, as demonstrated in our analytical models     

The Future of Simulation: A Field of Dreams

Due to the enormous complexity of computer systems, researchers use simulators to model system behavior and generate quantitative estimates of expected performance. Researchers also use simulators to model and assess the efficacy of future enhancements and novel systems. Arguably the most important tools available to computer architecture researchers, simulators offer a balance of cost, timeliness, and flexibility. Improving the infrastructure, benchmarking, and methodology of simulation - the dominant computer performance evaluation method - results in higher efficiency and let architects gain more insight into processor behavior. For these reasons, architecture researchers have increasingly relied on simulators