Reporting of Quantitative Protein Electrophoresis in Australia and New Zealand: a Call for Standardisation

Background

The lack of guidelines on reporting standards for protein electrophoresis may have led to significant differences in reports from different laboratories.

Objective

To determine the extent of variation in reporting of protein electrophoresis results in Australia and New Zealand.

Method

Questionnaires were distributed to laboratories throughout Australia and New Zealand asking about protein electrophoresis practices and reporting.

Results

Extensive variation was found in the following reporting practices: (a) units for urine Bence Jones protein (BJP); (b) reporting absence of a paraprotein rather than a normal pattern; (c) numerical reporting of all protein fractions or only the paraprotein; (d) warning of possible inaccuracy in the serum immunoglobulin result of the paraprotein type; (e) co-migration of a paraprotein with a normal serum protein; (f) use of a confirmatory test when a known paraprotein is no longer detectable.

Conclusions

A working party should be established to make recommendations on the reporting of protein electrophoresis. Implementation of such recommendations should reduce both report variation between laboratories and the risk of misinterpretation of reports.     

Rational design of a combination medication for the treatment of obesity

Existing obesity therapies are limited by safety concerns and modest efficacy reflecting a weight loss plateau. Here, we explore combination therapy with bupropion (BUP), a putative stimulator of melanocortin pathways, and an opioid antagonist, naltrexone (NAL), to antagonize an inhibitory feedback loop that limits sustained weight reduction. In vitro electrophysiologic experiments were conducted to determine the extent to which BUP+NAL stimulated hypothalamic pro-opiomelanocortin (POMC) neurons in mouse brain. A subsequent study further characterized the effect of combination BUP+NAL treatment on food intake in lean and obese mice. Finally, a randomized, blinded, placebo-controlled trial in obese adult subjects was conducted. Randomization included: BUP (300 mg) + NAL (50 mg), BUP (300 mg) + placebo (P), NAL (50 mg) + P or P+P for up to 24 weeks. BUP+NAL stimulated murine POMC neurons in vitro and caused a greater reduction in acute food intake than either monotherapy, an effect consistent with synergism. Combined BUP+NAL provided sustained weight loss without evidence of an efficacy plateau through 24 weeks of treatment. BUP+NAL completers diverged from NAL+P (P < 0.01) and P+P (P < 0.001) at week 16 and from BUP+P by week 24 (P < 0.05). The combination was also well tolerated. Translational studies indicated that BUP+NAL therapy produced synergistic weight loss which exceeded either BUP or NAL alone. These results supported the hypothesis that NAL, through blockade of beta-endorphin mediated POMC autoinhibition, prevents the classic weight loss plateau observed with monotherapies such as BUP. This novel treatment approach (BUP+NAL) holds promise for the treatment of obesity.\     

Effects of an exotic invasive macrophyte (tropical signalgrass) on native plant community composition,species richness and functional diversity

1. The issue of freshwater species being threatened by invasion has become central in conservation biology because inland waters exhibit the highest species richness per unit area, but apparently have the highest extinctions rates on the planet. 2. In this article, we evaluated the effects of an exotic, invasive aquatic grass (Urochloa subquadripara– tropical signalgrass) on the diversity and assemblage composition of native macrophytes in four Neotropical water bodies (two reservoirs and two lakes). Species cover was assessed in quadrats, and plant biomass was measured in further quadrats, located in sites where tropical signalgrass dominated (D quadrats) and sites where it was not dominant or entirely absent (ND quadrats). The effects of tropical signalgrass on macrophyte species richness, Shannon diversity and number of macrophyte life forms (a surrogate of functional richness) were assessed through regressions, and composition was assessed with a DCA. The effects of tropical signalgrass biomass on the likelihood of occurrence of specific macrophyte life forms were assessed through logistic regression. 3. Tropical signalgrass had a negative effect on macrophyte richness and Shannon and functional diversity, and also influenced assemblage composition. Emergent, rooted with floating stems and rooted submersed species were negatively affected by tropical signalgrass, while the occurrence of free‐floating species was positively affected. 4. Our results suggest that competition with emergent species and reduction of underwater radiation, which reduces the number of submersed species, counteract facilitation of free‐floating species, contributing to a decrease in plant diversity. In addition, homogenisation of plant assemblages shows that tropical signalgrass reduces the beta diversity in the macrophyte community. 5. Although our results were obtained at fine spatial scales, they are cause for concern because macrophytes are an important part of freshwater diversity.     

Analyzing complex traits with congenic strains

Congenic strains continue to be a fundamental resource for dissecting the genetic basis of complex traits. Traditionally, genetic variants (QTLs) that account for phenotypic variation in a panel of congenic strains are sought first by comparing phenotypes for each strain to the host (reference) strain, and then by examining the results to identify a common chromosome segment that provides the best match between genotype and phenotype across the panel. However, this “common-segment” method has significant limitations, including the subjective nature of the genetic model and an inability to deal formally with strain phenotypes that do not fit the model. We propose an alternative that we call “sequential” analysis and that is based on a unique principle of QTL analysis where each strain, corresponding to a single genotype, is tested individually for QTL effects rather than testing the congenic panel collectively for common effects across heterogeneous backgrounds. A minimum spanning tree, based on principles of graph theory, is used to determine the optimal sequence of strain comparisons. For two traits in two panels of congenic strains in mice, we compared results for the sequential method with the common-segment method as well as with two standard methods of QTL analysis, namely, interval mapping and multiple linear regression. The general utility of the sequential method was demonstrated with analysis of five additional traits in congenic panels from mice and rats. Sequential analysis rigorously resolved phenotypic heterogeneity among strains in the congenic panels and found QTLs that other methods failed to detect.     

长爪沙鼠线粒体DNA控制区全序列测定及分析

目的对长爪沙鼠线粒体DNA控制区全序列进行测定,并对其进行鉴定及进化分析。方法根据长爪沙鼠已知基因序列设计引物,采用PCR产物测序法,对所得的片段进行测序鉴定。结合已公布啮齿类动物D-loop区序列,分析其碱基组成、遗传距离、并基于最小进化法和UPGMA法构建系统进化树。结果获得长爪沙鼠D-loop区序列,其与家鼠、小家鼠和仓鼠平均同源性为58%;碱基组成分析显示,长爪沙鼠与啮齿类动物有相似的碱基组成和碱基偏离,其A-skew和G-skew分别为0.0047和-0.28。进化分析结果显示,长爪沙鼠与家鼠（0.35）、黑家鼠（0.38）和仓鼠（0.39）具有较近的遗传距离,其分化顺序为跳鼠、蔗鼠、长爪沙鼠、仓鼠、家鼠和小家鼠。结论本研究获得长爪沙鼠D-loop区全序列,确定了长爪沙鼠与仓鼠、家鼠、小家鼠及其它啮齿动物的进化关系,为长爪沙鼠进化研究、线粒体的结构和功能研究奠定基础。     

Cold adaptation in the marine bacterium,Sphingopyxis alaskensis,assessed using quantitative proteomics

The cold marine environment constitutes a large proportion of the Earth's biosphere. Sphingopyxis alaskensis was isolated as a numerically abundant bacterium from several cold marine locations, and has been extensively studied as a model marine bacterium. Recently, a metabolic labelling platform was developed to comprehensively identify and quantify proteins from S. alaskensis. The approach incorporated data normalization and statistical validation for the purpose of generating highly confident quantitative proteomics data. Using this approach, we determined quantitative differences between cells grown at 10°C (low temperature) and 30°C (high temperature). Cold adaptation was linked to specific aspects of gene expression: a dedicated protein‐folding system using GroESL, DnaK, DnaJ, GrpE, SecB, ClpB and PPIase; polyhydroxyalkanoate‐associated storage materials; a link between enzymes in fatty acid metabolism and energy generation; de novo synthesis of polyunsaturated fatty acids in the membrane and cell wall; inorganic phosphate ion transport by a phosphate import PstB homologue; TonB‐dependent receptor and bacterioferritin in iron homeostasis; histidine, tryptophan and proline amino acid metabolism; and a large number of proteins without annotated functions. This study provides a new level of understanding on how important marine bacteria can adapt to compete effectively in cold marine environments. This study is also a benchmark for comparative proteomic analyses with other important marine bacteria and other cold‐adapted organisms.     

Physiological evidence for the involvement of peptide YY in the regulation of energy homeostasis in humans

Objective: To explore the potential role of the endogenous peptide YY (PYY) in the long‐term regulation of body weight and energy homeostasis. Research Methods and Procedures: Fasting and postprandial plasma PYY concentrations were measured after an overnight fast and 30 to 180 minutes after a standardized meal in 29 (21 men/8 women) non‐diabetic subjects, 16 of whom had a follow‐up visit 10.8 ± 1.4 months later. Ratings of hunger and satiety were collected using visual analog scales. Resting metabolic rate (RMR) (15‐hour RMR) and respiratory quotient (RQ) were assessed using a respiratory chamber. Results: Fasting PYY concentrations were negatively correlated with various markers of adiposity and negatively associated with 15‐hour RMR (r = ?0.46, p = 0.01). Postprandial changes in PYY (area under the curve) were positively associated with postprandial changes in ratings of satiety (r = 0.47, p = 0.01). The maximal PYY concentrations achieved after the meal (peak PYY) were negatively associated with 24‐hour RQ (r = ?0.41, p = 0.03). Prospectively, the peak PYY concentrations were negatively associated with changes in body weight (r = ?0.58, p = 0.01). Discussion: Our data indicate that the endogenous PYY may be involved in the long‐term regulation of body weight. It seems that this long‐term effect was not exclusively driven by the modulation of food intake but also by the control of energy expenditure and lipid metabolism.     

To be or NUCB2, is nesfatin the answer?

Signals from the hypothalamus govern food intake and energy balance. A new study describes nesfatin-1, a hypothalamic and brainstem peptide whose expression decreases during fasting. Although central treatment with nesfatin-1 inhibited food intake and nesfatin-1 blockade increased food intake, the role and mechanism of nesfatin in energy balance remains unclear.     

Cell cycle related proteins in hyperplasia of usual type in breast specimens of patients with and without breast cancer

Background  

Hyperplasia of usual type (HUT) is a common proliferative lesion associated with a slight elevated risk for subsequent development of breast cancer. Cell cycle-related proteins would be helpful to determine the putative role of these markers in the process of mammary carcinogenesis. The aim of this study was to analyze the expression of cell cycle related proteins in HUT of breast specimens of patients with and without breast cancer, and compare this expression with areas of invasive carcinomas.