Biomarkers discovery in cancer--up-dates in methodology

Biomarkers are biomolecules that can indicate normal/pathological processes, or physiological responses to therapy. Due to the serum abundance in proteins, such as albumin and lypo/glycoproteins, biomarkers are difficult to assess. Serum biomarkers identification can contribute to personalized medicine and improve cancer diagnostic and prognostic. The paper summarizes some of the proteomics techniques and the workflow used for protein signatures identification associated to cancer development. Thus, biomarkers validated for prostatic, breast, cervical or lung cancers are presented as examples for clinical application of serum markers. In spite of the continuous research efforts, there are only few validated biomarkers that have proved a good predictive power in cancer. Modern technology and the combination of various techniques used for proteins quantification represent important means for the identification and validation of new biomarkers.     

Twinning in thoroughbred mares in Poland

Of the thoroughbred population bred in Poland from 1952 to 1976, an average of 3.3% twinnings were registered. Of these averages 73% resulted in abortion, 11% in still births, and 16% in the live birth of one or both of the foals. Twins were not any more valuable for racing or breeding than normal. Mares mated in June had a twinning coefficient of 6.6% and those mated from February to May had one from 3.0% to 3.6%. The twinning coefficient was higher for mares which had not lactated during the previous year (6.0%, p 相似文献   

Problems in maintenance therapy in acute myeloid leukemias in adults]

Problems of maintaining therapy for acute myelocytic leukemias in adults are discussed. The analysis of the maintaining therapy in 22 patients affected with an acute myelocytic leukemia and living for more than 6 months revealed that the interval therapy with a high dosage of cytostatic combinations in the sense of the COAP scheme is preferable compared with the daily administration of 6-mercaptopurin, in addition methotrexate twice a week. Reasons for this are discussed.     

The NIH-shift in the in vivo hydroxylation of ring-deuterated L-phenylalanine in man

Oral loading with L-[ring-2H5]phenylalanine has been performed at a dose of 25 mg/kg for detection of heterozygotes for classic phenylketonuria. Using three differently labeled batches of ring-deuterated L-phenylalanine, quantitative analysis of deuterium-labeled L-phenylalanine and L-tyrosine in plasma revealed different label distributions. Three different reaction mechanisms for the 4-hydroxylation of L-phenylalanine to L-tyrosine were used as the basis for model calculations of the transformation of the L-phenylalanine label distribution into that of L-tyrosine. The best agreement between observed and calculated distributions was found for the mechanism involving a migration of the 4-substituent into the 3- or 5-position (NIH-shift), followed by a random loss of the 4-/3- or the 4-/5-substituent from this intermediate structure.     

Changes in ventilation in response to ramp changes in treadmill exercise load

These experiments examined the changes in ventilation during a 40-s ramp increase in exercise load, produced by increasing either the speed of the treadmill or the grade, to equivalent end-points of oxygen uptake. Six subjects underwent five trials each for grade and speed, while ventilation was monitored breath-by-breath. For each subject, ventilation versus time for all five of the speed trials was plotted on a single graph and fitted by linear regression. The data for the grade trials were similarly treated. For all subjects, the slope of the regression line for the speed plots was found to be significantly (P < 0.05) greater than that for the grade plots. We concluded that these experimental results support the hypothesis that the neural drive to ventilation persists as exercise continues and is proportionately related to the frequency of limb movement.     

Diversification of exogenous genes in vivo in Neurospora

We have adapted the meiotic recombination hotspot cog of Neurospora crassa for shuffling exogenous DNA, providing a means of generating novel genes in situ from sequences introduced into chromosomes. Genes to be diversified are inserted between the his-3 locus and cog. Diversification crosses are heterozygous both for alleles of the exogenous DNA and for auxotrophic alleles of his-3. Progeny selected for ability to grow without histidine supplementation are enriched for exchange events within the exogenous DNA. Exchange events initiated by cog can propagate past DNA sequences mismatched for more than 370 bp and complete exchanges in patches of matched sequence as short as 24 bp, parameters that make the system suited for use in the directed evolution of genes for protein engineering. Here we demonstrate the system by shuffling human immunoglobulin kappa chain genes and also endoglucanase genes derived from different species of fungi.     

D-Methylphenidate is non-genotoxic in in vitro and in vivo assays

D-Methylphenidate (dexmethylphenidate; D-MPH) and its racemate D,L-methylphenidate (D,L-MPH) are currently prescribed for the chronic treatment of attention deficit hyperactivity disorder (ADHD) in children. Studies have shown that D-MPH is the pharmacologically active enantiomer for ADHD and is therefore the preferred drug for the treatment of ADHD symptoms. Although studies on the mutagenicity of D,L-MPH have been conducted, similar data for D-MPH are lacking. Therefore, D-MPH was evaluated in the bacterial reverse mutation and mouse lymphoma assays with and without S9 and in a bone marrow micronucleus test in male and female CD-1 mice. As a comparison, the L-enantiomer and racemate were also included in the assessments. While MPH-associated toxicity was observed in the mammalian tests, none of the three compounds tested induced mutagenic or clastogenic effects. Our present results along with published epidemiological data from patient populations are consistent with the conclusion that D-MPH and D,L-MPH do not present a carcinogenic risk to humans.     

Meiosis in Bovine Oocytes Matured in Vitro and in Vivo

Meiosis in bovine oocytes has; been studied after maturation in vitro or in vivo. Oocytes for in vitro maturation were collected from the ovaries of slaughtered cattle without regard to the phase of the estrous cycle while in vivo maturation was studied in oocytes from gonadotrophin-stimulated heifers at times varying between 6 and 36 h after the beginning of behavioural estrus. Oocytes from slaughtered cattle were classified according to their cumulus complex and ooplasm and were cultured for 6, 12, 18, 24, 36 or 48 h in modified Krebs-Ringer bicarbonate buffer before fixation) for cytogenetic analysis. Oocytes from stimulated heifers were aspirated from follicles or flushed from the oviducts, classified according to cumulus and ooplasm, and fixed within 6 h of collection. Nuclear maturation was more rapid in vitro than in vivo. The largest proportion of oocytes reached maturity (Mil) after 12 to 18 h in culture or 30 to 36 h after the onset of behavioural estrus. Oocytes devoid of cumulus cells or showing signs of vacuolation or degeneration had virtually no capacity for nuclear maturation.     

Improvements in cytological preparations for fluorescent in situ hybridization in Passiflora

Cytological preparations for the fluorescent in situ hybridization (FISH) technique require cytoplasm-free metaphases, with well-spread chromosomes, for the localization of DNA sequences and chromosome mapping. We tested various procedures for FISH analysis of Passiflora cacaoensis, P. gardneri and hybrid F? progeny of P. gardneri x P. gibertii. Two treatments with four enzymes and three incubation times were compared. The material was treated with 1.0 M HCl before enzymatic digestion. The following criteria were used to determine the quality of the metaphases: a) lack or presence of cytoplasm; b) well-spread chromosomes or with overlap; c) complete or incomplete chromosome number (2n). The enzyme Pectinex(?) SP ULTRA gave the best performance, with the shortest incubation time. The best results were observed after 30 min of incubation; more than 70% of the metaphases did not have large amounts of cytoplasm or overlapping chromosomes, and about 75% maintained the chromosome number. FISH was carried out using a 45S rDNA probe (pTa71) labeled with biotin and detected with fluorescein isothiocyanate. Sites with strong staining and without nonspecific signals were observed. Our methodological adaptations allowed the preparation of metaphase slides of high quality for the FISH technique, with less time required for the preparation of samples.     

Recent advances in Pelargonium in vitro regeneration systems

Recent advances in the development of protocols for in vitro culture and genetic manipulation have provided new avenues for the development of novel varieties of Pelargonium and for use as model systems for investigating the factors controlling plant morphogenesis. Optimized techniques of meristem culture have supplemented the culture indexing methods in commercial greenhouse production resulting in availability of large-scale pathogen indexed planting material. Currently, technologies are available for the mass in vitro propagation of F1 hybrid Pelargonium through both organogenesis and somatic embryogenesis. The somatic embryogenesis model system has allowed researchers to identify critical factors controlling plant morphogenesis in vitro such as regulation of regeneration by growth regulators, choice of explant and characterization of induction and expression phases of morphogenesis in Pelargonium. Also, optimization of technologies for genetic transformation of Pelargonium opened up the possibilities for developing genotypes with novel characters, including resistance to some of the major diseases. Finally, the development of regeneration systems for Pelargonium spp. has facilitated conventional crop improvement programs, thereby providing a valuable resource to the horticultural industry.     

Specific features of in vivo and in vitro sperm storage in birds

This review focuses on some of the main features of sperm selection and storage in birds mainly on the basis of studies performed in poultry species, with emphasis on the initial selection of sperm at the female vagina level prior to migration towards the sperm storage tubules. Sperm originating from low-quality males or subjected to inappropriate in vitro storage conditions are rapidly discarded, resulting in impaired fertility in corresponding flocks. In the absence of accessible and appropriate technology for matching the 'storing' potential of sperm in the oviduct, conditions for prolonged sperm storage under a liquid (through the use of semen extenders) or a solid state (cryopreservation) have received only limited attention, despite their potential interest to facilitate male and female management in poultry flocks. Despite this, technology for short-term liquid storage is currently used in turkeys, guinea fowl and muscovy ducks and also in progress in chickens. In addition, technology for cryopreservation of avian semen has become available for some species (chicken, goose) to facilitate the management of genetic resources, including the preservation of rare and economically important breeds.     

二型态真菌形态调控研究现状与展望

真菌二型态(Dimorphism)是指某些真菌在外界环境因子的诱导下,其营养体可在酵母型(Yeast form)和菌丝型(Mycelium form)两种不同细胞形态间转化的能力。诱导二型态真菌形态转化的环境因子众多,不同环境因子诱导二型态转化的分子机制也不尽相同。二型态真菌形态转换常与致病性有关,在微生物发酵调控中也至关重要,研究真菌形态调控不仅有助于阐明二型态真菌的致病机理,还可丰富二型态真菌形态调控在发酵工业中的应用。本文从影响真菌二型态的外界环境因子、转化机制和真菌发酵形态控制等方面对二型态真菌形态调控研究现状及应用前景进行综述。     

ADHD in adults: a concept in evolution

Although attention-deficit/hyperactivity disorder (ADHD) has been recognized as a disorder affecting individuals across the life cycle since the end of the nineties, there is still considerable debate on how to conceptualize the disorder in adults, and on the best way to operationalize diagnostic criteria for this age range. In this comprehensive non-systematic review of the literature, we provide data about prevalence and presentation of ADHD in adulthood as well as discuss major problems in applying criteria developed for children in assessing adults (clinical utility, threshold of symptoms for diagnosis, full ADHD diagnosis in childhood, information source, and additional dimensions for diagnosis-executive functioning impairment and emotional impulsivity). In addition, we provide some recommendations for improving ADHD diagnostic criteria in adulthood.     

Seven-Base-Pair Inverted Repeats in DNA Form Stable Hairpins in Vivo in Saccharomyces Cerevisiae

Palindromic sequences in single-stranded DNA and RNA have the potential for intrastrand base pairing, resulting in formation of "hairpin" structures. We previously reported a genetic method for detecting such structures in vivo in the yeast Saccharomyces cerevisiae. Below, we describe evidence indicating that a 14-base-pair palindrome (7 bp per inverted repeat) is sufficient for formation of a hairpin in vivo.     

A survey of avian influenza in tree sparrows in China in 2011

Tree sparrows (Passer montanus) are widely distributed in all seasons in many countries. In this study, a survey and relevant experiments on avian influenza (AI) in tree sparrows were conducted. The results suggested that the receptor for avian influenza viruses (AIVs), SAα2,3Gal, is abundant in the respiratory tract of tree sparrows, and most of the tree sparrows infected experimentally with two H5 subtype highly pathogenic avian influenza (HPAI) viruses died within five days after inoculation. Furthermore, no AIVs were isolated from the rectum eluate of 1300 tree sparrows, but 94 serological positives of AI were found in 800 tree sparrows. The serological positives were more prevalent for H5 subtype HPAI (94/800) than for H7 subtype AI (0/800), more prevalent for clade 2.3.2.1 H5 subtype HPAI (89/800) than for clade 2.3.4 (1/800) and clade 7.2 (4/800) H5 subtype HPAI, more prevalent for clade 2.3.2.1 H5 subtype HPAI in a city in southern China (82/800) than in a city in northern China (8/800). The serological data are all consistent with the distribution of the subtypes or clades of AI in poultry in China. Previously, sparrows or other passerine birds were often found to be pathogenically negative for AIVs, except when an AIV was circulating in the local poultry, or the tested passerine birds were from a region near waterfowl-rich bodies of water. Taken together, the data suggest that tree sparrows are susceptible to infection of AIVs, and surveys targeting sparrows can provide good serological data about the circulation of AIVs in relevant regions.     

Variations in WGA-binding proteins in tumours growing in different implantation sites

The protein/glycoprotein composition of Triton X100 extracts of four metastatic tumour lines have been compared by gel electrophoresis after the implantation into different sites of the body. No consistent differences in the patterns were noted after staining with Coomassie Blue or treating with concanavalin A. In contrast, after treating with wheat germ agglutinin (WGA), the patterns varied with site. This was most marked with the ML and M5 tumours, where the overall binding was much lower for some sites. Although WGA binding with the LL and BL6 tumours was generally higher than with the ML and M5, reduced binding was still evident for some sites. Reduced WGA binding was also observed for extracts of secondary tumour growing in the liver or lungs.     

Fluorescent in situ visualization of sterols in Arabidopsis roots

Sterols are eukaryotic membrane components with crucial roles in diverse cellular processes. Elucidation of sterol function relies on development of tools for in situ sterol visualization. Here we describe protocols for in situ sterol localization in Arabidopsis thaliana root cells, using filipin as a specific probe for detection of fluorescent filipin-sterol complexes. Currently, filipin is the only established tool for sterol visualization in plants. Filipin labeling can be performed on aldehyde-fixed samples, largely preserving fluorescent proteins and being compatible with immunocytochemistry. Filipin can also be applied for probing live cells, taking into account the fact that it inhibits sterol-dependent endocytosis. The experimental procedures described are designed for fluorescence detection by confocal laser-scanning microscopy with excitation of filipin-sterol complexes at 364 nm. The protocols require 1 d for sterol covisualization with fluorescent proteins in fixed or live roots and 2 d for immunocytochemistry on whole-mount roots.     

中国食蚜蝇科昆虫研究进展

食蚜蝇Syrphidae是双翅目昆虫中一类很重要的资源昆虫和天敌昆虫,在控制蚜虫、自然界传粉等方面有很重要的作用,同时也是仿生学等研究的重要实验材料。文章介绍国内食蚜蝇科昆虫最近几年的分类学、生态学、生物学习性及生物防治的研究现状。     

Novel proteins in emulsions using in vitro compartmentalization

IVC (in vitro compartmentalization) provides a complete cell-free approach for the production of novel targeted proteins. IVC uses aqueous droplets, which contain DNA and components for protein production, within water-in-oil emulsions. Recent advances in the composition and formation, as well as the detection, sorting and recovery, of the droplets enable the evolution of the encoded protein. Furthermore, IVC technology permits the step-wise addition of reagents into the droplets, making them suitable for high-throughput applications - where synthetic enzymes with substrate specificity are selected for catalytic activity, binding and regulation. In the broad field of in vitro display, developments such as the incorporation of unnatural amino acids and the production of cell toxic proteins expand the diverse spectrum of future applications for IVC.