<Emphasis Type="Italic">Streptococcus agalactiae</Emphasis> CAMP factor/protein B does not bind to human IgG

CAMP factor is an extracellular cytolytic protein produced by Streptococcus agalactiae. CAMP factor has been reported to bind the Fc fragments of immunoglobulin G (IgG) and has therefore also been called protein B, in analogy to protein A of Staphylococcus aureus. We attempted to characterize the interaction of protein B with IgG in more detail. In contrast to protein A, CAMP factor does not inhibit the activation of complement by hemolysin antibodies bound to sheep red cell surfaces. IgG also failed to inhibit the co-hemolytic activity of CAMP factor, which is in disagreement with previous findings. After co-incubation, CAMP factor and IgG were cleanly separated by gel filtration, indicating that no binding had occurred. Waseem El-Huneidi and Ryan Mui contributed equally to this work.     

Protein Interfacial Pocket Engineering via Coupled Computational Filtering and Biological Focusing Criterion

To engineer bio-macromolecular systems, protein–substrate interactions and their configurations need to be understood, harnessed, and utilized. Due to the inherent large numbers of combinatorial configurations and conformational complexity, methods that rely on heuristics or stochastics, such as practical computational filtering (CF) or biological focusing (BF) criterions, when used alone rarely yield insights into these complexes or successes in (re)designing them. Here we use a coupled CF–BF criterion upon an amenable interfacial pocket (IP) of a protein scaffold complexed with its substrate to undergo residue replacement and R-group refinement (R⁴) to filter out energetically unfavorable residues and R-group conformations, and focus in on those that are evolutionarily favorable. We show that this coupled filtering and focusing can efficiently provide a putative engineered IP candidate and validate it computationally and empirically. The CF–BF criterion may permit holistic understanding of the nuances of existing protein IPs and their scaffolds and facilitate bioengineering efforts to alter substrate specificity. Such approach may contribute to accelerated elucidation of engineering principles of bio-macromolecular systems. Electronic supplementary material The online version of this article (doi: ) contains supplementary material, which is available to authorized users.     

In vivo immunostimulatory effects of CpG ODN in newborn piglets

The in vivo immunoadjuvant effects of CpG oligodeoxynucleotides (CpG ODN) have been studied extensively in mice and relatively fewer studies have been done in other species. But so far, the innate immunostimulatory effects of CpG ODN have been demonstrated just in mouse, monkey, sheep and chicken in some reports. The purpose of this study is to determine the potential effects of CpG ODN in newborn piglets. The proportion of CD4(+), CD8(+) T lymphocytes subpopulations and the major histocompability complex (MHC-II) antigen expression of peripheral blood mononuclear cells (PBMCs) and IFN-gamma in serum were tested at various time-points. The results suggested that, the CD4(+)/CD8(+) ratio decreased over time in piglets inoculated with phosphate buffer saline (PBS) alone, however, it was stable in CpG ODN-inoculated piglets; the use of CpG ODN can prevent effectively the reduction of the proportion of CD4(+) T lymphocytes. The MHC-II antigen expression and IFN-gamma level of CpG ODN-injected piglets were significantly higher than those of PBS-injected piglets. The ODN-induced responses were stronger in animals injected with CpG ODN formulated in 30% emulsigen than in PBS. The innate immunostimulatory activity of CpG ODN appeared to be in dose-dependent manner. These in vivo data demonstrate for the first time that CpG ODN can stimulate innate immune system in newborn piglets.     

Neuroprotective effects of safflor yellow B on brain ischemic injury

The present study was conducted to investigate whether safflor yellow B (SYB) had a protective effect on cerebral ischemic injury and to determine the possible mechanisms in vivo and in vitro. In vivo, Male Wistar–Kyoto (WKY) rats were used to make the model of middle cerebral artery occlusion (MCAO). The behavioral test was used to measure neurological deficit scores for evaluation of the ischemic damage of brain. The infarction area of brain was assessed in brain slices stained with 2% solution of 2,3,5-triphenyl tetrazolium chloride (TTC). Spectrophotometric assay was used to determine the activities of superoxide dismutase (SOD) and glutathione peroxidase (GPx), contents of malondialdehyde (MDA) and adenosine triphosphate (ATP) of the brain. Furthermore, the respiratory control ratio (RCR = state 3/state 4) was assessed in the brain mitochondria. In vitro, the effect of SYB was tested in cultured fetal cortical cells exposed to glutamate to identify its neuroprotection against neurons damage. The results in vivo showed that SYB at doses of 3.0 and 6.0 mg kg⁻¹ markedly decreased the neurological deficit scores and the infarction area in MCAO rats. At the same time, SYB significantly improved mitochondrial energy metabolism, decreased MDA content, and increased SOD and GPx activities in ischemic brain. The results in vitro showed that SYB remarkably inhibited neuron damage induced by glutamate in cultured fetal cortical cells. These suggest that SYB might act as a potential neuroprotective agent against the cerebral ischemia-induced injury in rat brain through reducing lipid peroxides, scavenging free radicals, and improving the energy metabolism.     

Temporal dynamics of semicircular canal and otolith function following acute unilateral vestibular deafferentation in humans

Dynamic changes of deficits in canal and otolith vestibulo-ocular reflexes (VORs) to high acceleration, eccentric yaw rotations were investigated in five subjects aged 25–65 years before and at frequent intervals 3–451 days following unilateral vestibular deafferentation (UVD) due to labyrinthectomy or vestibular neurectomy. Eye and head movements were recorded using magnetic search coils during transients of directionally random, whole-body rotation in darkness at peak acceleration 2,800°/s². Canal VORs were characterized during rotation about a mid-otolith axis, viewing a target 500 cm distant until rotation onset in darkness. Otolith VOR responses were characterized by the increase in VOR gain during identical rotation about an axis 13 cm posterior to the otoliths, initially viewing a target 15 cm distant. Pre-UVD canal gain was directionally symmetrical, averaging 0.87 ± 0.02 (±SEM). Contralesional canal gain declined from pre-UVD by an average of 22% in the first 3–5 days post-UVD, before recovering to an asymptote of close 90% of pre-UVD level at 1–3 months. This recovery corresponded to resolution of spontaneous nystagmus. Ipsilesional gain declined to 59%, and showed no consistent recovery afterwards. Pre-UVD otolith gain was directionally symmetrical, averaging 0.56 ± 0.02. Immediately after UVD, the contralesional otolith gain declined to 0.30 ± 0.02, and did not recover. Ipsilesional otolith gain declined profoundly to 0.08 ± 0.03 (P < 0.01), and never recovered. In contrast to the modest and directionally symmetrical effect of UVD on the human otolith VOR during pure translational acceleration, otolith gain during eccentric yaw rotation exhibited a profound and lasting deficit that might be diagnostically useful in lateralizing otolith pathology. Most recovery of the human canal gain to high acceleration transients following UVD is for contralesional head rotation, occurring within 3 months as spontaneous nystagmus resolves. Grant support: United States Public Health Service grants DC-02952 and AG-09693. JLD is Leonard Apt Professor of Ophthalmology.     

Protein tyrosine phosphatase 1B inhibitors from natural sources

Since PTP1B enzyme was discovered in 1988, it has captured the research community’s attention. This landmark discovery has stimulated numerous research studies on a variety of human diseases, including cancer, inflammation, and diabetes. Tremendous progress has been made in finding PTP1B inhibitors and exploring PTP1B regulatory mechanisms. This review investigates for the natural PTP1B inhibitors, and focuses on the common characteristics of the discovered structures and structure–activity relationships. To facilitate understanding, all the natural compounds are here divided into five different classes (fatty acids, phenolics, terpenoids, steroids, and alkaloids), according to their skeletons. These PTP1B inhibitors of scaffold structures could serve as a theoretical basis for new concept drug discovery and design.     

Contribution of equilibrative nucleoside transporters 1 and 2 to gemcitabine uptake in pancreatic cancer cells

Hepatic arterial infusion (HAI) chemotherapy is expected to be a more effective and safer method to treat the hepatic metastasis of pancreatic cancer than intravenous (iv) administration because of higher tumor exposure and lower systemic exposure. To clarify the uptake mechanism of nucleoside anticancer drugs, including gemcitabine (GEM), in pancreatic cancer, we investigated the uptakes of radiolabeled uridine (a general substrate of nucleoside transporters) and GEM in pancreatic cancer cell lines MIA‐PaCa2 and As‐PC1. Uridine uptake was inhibited by non‐labeled GEM and also by S‐(4‐nitrobenzyl)‐6‐thioinosine (NBMPR; an inhibitor of equilibrative nucleoside transporters, ENTs) in a concentration‐dependent manner, suggesting that ENTs contribute to uridine uptake in pancreatic cancer cells. As for GEM, saturable uptake was mediated by high‐ and low‐affinity components with K_m values of micromolar and millimolar orders, respectively. Uptake was inhibited in a concentration‐dependent manner by NBMPR and was sodium ion‐independent. Moreover, the concentration dependence of uptake in the presence of 0.1 μM NBMPR showed a single low‐affinity site. These results indicated that the high‐ and low‐affinity sites correspond to hENT1 and hENT2, respectively. The results indicated that at clinically relevant hepatic concentrations of GEM in GEM‐HAI therapy, the metastatic tumor exposure of GEM is predominantly determined by hENT2 under unsaturated conditions, suggesting that hENT2 expression in metastatic tumor would be a candidate biomarker for indicating anticancer therapy with GEM‐HAI.     

贫困、疾病及精准健康扶贫政策:基于贫困居民生命质量评价

目的:将生命质量评价引入精准健康识别与干预政策中,通过比较不同类别贫困居民生命质量特征,揭示扶贫对象各维度现状,为完善精准健康扶贫政策提供参考。方法:评价工具采用WHOQOL-BREF量表,通过入户调查方式收集数据,采用t检验分析与中国常模数据进行比较。结果:贫困居民其人群分布表现为男性(71.7%)多于女性,年龄普遍偏大(平均55岁)。贫困居民与中国常模比较,健康组和疾病组的生命质量评价评分均显著低于中国常模的对照组,说明贫困对于居民生命质量评价的负面影响十分显著。不论处于何种收入水平,健康贫困居民总体评价均高于患病贫困居民。对于患病贫困居民而言,收入越高、其心理和总体评价越好;反之亦然。结论:健康对于贫困居民总体评价具有显著正影响、而患病对于贫困居民总体评价具有显著负影响。收入提高对于患病贫困居民的心理和总体评价改善有效。