Cathodoluminescence Microscopy: A Useful Tool for Assessing Incremental Chemical Variation in Otoliths

Otoliths taken from fish from Eden Lake, Manitoba show yellow–green and red cathodoluminescence of varying intensity that corresponds to their annular structure. Proton-induced X-ray emission analysis shows manganese (Mn) concentrations of between 2 and 205ppm, zinc (Zn) concentrations between 2 and 290ppm and strontium (Sr) concentrations up to 1500ppm in the otoliths. The distribution of luminescence correlates with the distribution of Mn. The Mn, Zn and Sr are likely derived from the monzonitic rocks surrounding the lake. Variations in the distribution of cathodoluminescence may be a useful tool for evaluating changes in environmental chemistry and fish life histories.     

Solid-phase Chemistry: A Useful Tool to Discover Modulators of Protein Interactions

The Solid phase synthesis (SPS) concept, first developed for biopolymers, has spread in every field where organic synthesis is involved. While the potential of the solid-phase method was obvious in 1959 to its discoverer, Prof. R. B. Merrifield, it was unpredictable its dominance in peptide synthesis and especially in combinatorial chemistry, an area not yet conceived. SPS paved the way for solid-phase combinatorial approaches (extensively reviewed in (Choong, I. C. and Ellman, J. A.: 1996, Annu. Rep. Med. Chem. 31, 309–318; Obrecht, D. and Villalgordo, J. M.: 1998, Solid-supported Combinatorial and Parallel Synthesis of Small-Molecular-Weight Compound Libraries. Pergamon Press Ltd., Oxford, UK; Chabala, J. C.: 1995, Curr. Opin. Biotechnol. 6, 632–639; Kamal, A., Reddy, K. L., Devaiah, V., Shankaraiah, N., Reddy, D. R.: 2006, Mini Rev. Med. Chem. 6, 53–69; Whitehead, D. M., McKeown, S. C., Routledge, A.: 2005, Comb. Chem. HTS 8, 361–371; Nefzi, A., Ostresh, J. M., Houghten, R. A.: 1997, Chem. Rev. 97, 449–472; Gordon, E. M., Gallop, M. A., Patel, D. V.: 1996, Acc. Chem. Res. 29, 144–154)) as many laboratories and companies focused on the development of technologies and chemistry suitable to this new methodology. This resulted in the spectacular outburst of combinatorial chemistry, which profoundly changed the approach for new drug discovery. Combinatorial chemistry is currently considered a valid approach for a wide range of biomedical applications, such as, target validation and drug discovery.     

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of the dendritic tree, which eventually leads to neuronal output of action potentials at the axon. The influence of diverse spatial and temporal parameters of specific synaptic input on neuronal output is currently under investigation, e.g. the distance-dependent attenuation of dendritic inputs, the location-dependent interaction of spatially segregated inputs, the influence of GABAergig inhibition on excitatory integration, linear and non-linear integration modes, and many more.With fast micro-iontophoresis of glutamate and GABA it is possible to precisely investigate the spatial and temporal integration of glutamatergic excitation and GABAergic inhibition. Critical technical requirements are either a triggered fluorescent lamp, light-emitting diode (LED), or a two-photon scanning microscope to visualize dendritic branches without introducing significant photo-damage of the tissue. Furthermore, it is very important to have a micro-iontophoresis amplifier that allows for fast capacitance compensation of high resistance pipettes. Another crucial point is that no transmitter is involuntarily released by the pipette during the experiment.Once established, this technique will give reliable and reproducible signals with a high neurotransmitter and location specificity. Compared to glutamate and GABA uncaging, fast iontophoresis allows using both transmitters at the same time but at very distant locations without limitation to the field of view. There are also advantages compared to focal electrical stimulation of axons: with micro-iontophoresis the location of the input site is definitely known and it is sure that only the neurotransmitter of interest is released. However it has to be considered that with micro-iontophoresis only the postsynapse is activated and presynaptic aspects of neurotransmitter release are not resolved. In this article we demonstrate how to set up micro-iontophoresis in brain slice experiments.     

A Useful Accessory to the Zeiss Mechanical Stage for Oil Immersion Microscopy

Crystallia ribonuclease has no marked proteolytic activity, since digestion of sections with this enzyme produces no appreciable decrease in the intensity of the cytochemical tests for arginine and tyrosine. Cytoplasmic basophilia is unaffected by treatment with cold trichloracetic acid or with boiling alcohol-ether mixture. Mononucleotides and fatty acids thus have nothing to do with basophilia. Digestion of sections with desoryribonuclease has no effect on the alkaline phosphatase or arginine tests, while it supresses the Feulgen reaction and the affinity of the chromatin for basic and for some acid dyes.     

Self-Esteem and Obesity in Children and Adolescents: A Literature Review

The relationship between self-esteem and obesity has not received a great deal of empirical evaluation using strong research methodologies. Thus, it is not clear whether self-esteem is consistently related to obesity, whether the relationship is global or specific to physical appearance, whether the relationship differs by demographic variables such as age, gender or race/ethnicity, or whether self-esteem moderates weight changes during weight loss treatment programs. This review examines these questions using empirical evidence from 35 studies on the relationship between self-esteem and obesity in children and adolescents. Thirteen of 25 cross sectional studies clearly showed lower self-esteem in obese adolescents and children. Five of the six cross-sectional studies that included a measure of body esteem found lower body esteem in obese compared to normal weight children and adolescents. Results from two prospective studies examining initial self-esteem and later obesity were inconsistent. Results from six of eight treatment studies showed that weight loss treatment programs appear to improve self-esteem. However, it is unclear whether increases in self-esteem are related to enhanced weight loss. Many studies were methodologically weak primarily due to small and select samples and lack of appropriate comparison groups. Implications for prevention and treatment of childhood obesity are discussed.     

Peripheral Blood Lymphocyte Typing as a Useful Tool to Objectify the Oral Mucosa Patch Test in the Diagnosis of Allergic Contact Mucositis to Nickel

Nickel (Ni) exposure through the intestinal mucosa may cause a hypersensitivity reaction recently defined as allergic contact mucositis (ACM). This condition is identifiable by the oral mucosa patch test (omPT), a qualitative and subjective examination that requires clinical expertise. Our aim was to evaluate if a peripheral blood lymphocyte typing performed before and after the omPT for Ni may be able to objectify this examination for diagnostic purposes. Thirty patients with symptoms referable to the ingestion of Ni-rich foods were subjected to omPT for Ni. Before and after the omPT, each patient underwent blood sampling for the typing of total lymphocytes and their subsets (T, T helper or Th, T cytotoxic or Tc, B, natural killer or NK). Statistical analysis was performed by Student t test and receiver operating characteristic (ROC) curve analysis. According to the omPT outcomes, 18 patients were defined as Ni-sensitive and the remaining 12 as controls. In Ni-sensitive patients, the number of total, T, Th, Tc, and B lymphocytes/μL whole blood increased after the omPT (p?<?0.0001 for the first three, p?=?0.0004 and p?=?0.0001 for the last two lymphocyte types). No omPT-dependent lymphocyte increase was observed in controls. The post/pre omPT cell ratio, especially if calculated for Th lymphocytes, appears to be an effective index for diagnostic purposes (sensitivity?=?100 %, specificity?=?83.3 %, Youden index?=?0.833, area under curve (AUC)?=?0.926, p?<?0.0001). In conclusion, the peripheral blood lymphocyte typing with calculation of post/pre omPT cell ratio has the potential to support the omPT in diagnosing ACM, with the advantage of providing quantitative and objective data.     

Trends in Overweight and Obesity among Children and Adolescents in China from 1981 to 2010: A Meta-Analysis

Background

Overweight/obesity is a serious public health problem that affects a large part of the world population across all age and racial/ethnic groups. However, there has not been a meta-analysis of the prevalence of childhood and adolescent overweight/obesity in China during the past 30 years.

Methods

The China National Knowledge Infrastructure and Wanfang DATA, MEDLINE, EMBASE and Cumulative Index to Nursing and Allied Health Literature were searched for relevant studies published between January 1970 and June 2012. The prevalence of overweight/obesity over time was pooled using Stata/SE, version 9. Summary statistics (odds ratios, ORs) were used to compare sex-specific and urban-rural preponderance of overweight/obesity using Review Manager.

Results

After screening 1326 papers, we included 35 papers (41 studies), most of medium quality. The prevalence of overweight/obesity increased from 1.8% (95% confidence interval [CI], 0.4%–3.1%) and 0.4% (95% CI, −0.1% to −0.8%) respectively in 1981–1985 to 13.1% (95% CI, 11.2%–15.0%) and 7.5% (95% CI, 6.6%–8.4%) respectively in 2006–2010. The average annual increase was 8.3% and 12.4% respectively. Boys were more likely to be overweight/obese than girls (OR, 1.36; 95% CI, 1.24–1.49 and OR, 1.68; 95% CI, 1.52–1.86 respectively). The prevalence of overweight/obesity was higher in urban areas than in rural areas (OR, 1.66; 95% CI, 1.54–1.79 and OR, 1.97; 95% CI, 1.68–2.30 respectively). For age-specific subgroup analyses, both overweight and obesity increased more rapidly in the toddler stage than in other developmental stages. Sensitivity analyses showed that sample-size differences, study quality, overweight/obesity criteria and geographical distribution affected overweight/obesity prevalence.

Conclusions

Toddlers and urban boys were at particularly high risk; the prevalence in these groups increased more rapidly than in their counterparts. Public health prevention strategies are urgently needed to modify health behaviors of children and adolescents and control overweight/obesity in China.     

The Influence of Hand Preference on Grip Strength in Children and Adolescents; A Cross-Sectional Study of 2284 Children and Adolescents

Introduction

In adults the preferred hand is often considered to be around 10% stronger than the non-preferred hand. Whether the same is true for children and adolescents remains unclear. The objective of this study is therefore to determine whether there is a difference in grip strength between the preferred and non-preferred hand in developing children, to establish whether this difference is similar for children of a different gender or hand preference, and whether there is a difference in grip strength of the preferred hand of left-preferent (LP) and right-preferent (RP) children.

Design

Cross-sectional study.

Participants

Participants were recruited from schools in the northern provinces of the Netherlands. The study included healthy children and adolescents in the age range of 4–17 years.

Outcome Measures

Each child was allowed a total of four attempts using the JAMAR hand dynamometer, two attempts with each hand. All individual attempts were scored. Hand preference was determined by asking which hand was used to write, or in the case of 4- and 5-year-olds, which hand was used to draw a shape.

Results

The study population comprised 2284 children and adolescents. RP boys and girls scored significantly higher with their preferred hand, the difference amounting to 9.5 and 10.1% respectively. LP girls scored significantly higher with their preferred hand, but this difference was only 3.0%. For LP boys no significant difference was found in favor of either hand. LP children score higher with the non-preferred hand and tie scores on both hands more often than RP children.

Conclusion

The 10% rule of hand preference is applicable to RP children ranging in age between 4 and 17 years, but not to LP children. In contrast to LP boys, LP girls are generally significantly stronger with their preferred hand.     

Cancer Risk in Children and Adolescents with Birth Defects: A Population-Based Cohort Study

Objective

Birth defects are an increasing health priority worldwide, and the subject of a major 2010 World Health Assembly Resolution. Excess cancer risk may be an added burden in this vulnerable group of children, but studies to date have provided inconsistent findings. This study assessed the risk for cancer in children and young adolescents with major birth defects.

Methods and Findings

This retrospective, statewide, population-based, cohort study was conducted in three US states (Utah, Arizona, Iowa). A cohort of 44,151 children and young adolescents (0 through 14 years of age) with selected major, non-chromosomal birth defects or chromosomal anomalies was compared to a reference cohort of 147,940 children without birth defects randomly sampled from each state’s births and frequency matched by year of birth. The primary outcome was rate of cancer prior to age 15 years, by type of cancer and type of birth defect. The incidence of cancer was increased 2.9-fold (95% CI, 2.3 to 3.7) in children with birth defects (123 cases of cancer) compared to the reference cohort; the incidence rates were 33.8 and 11.7 per 100,000 person-years, respectively. However, the excess risk varied markedly by type of birth defect. Increased risks were seen in children with microcephaly, cleft palate, and selected eye, cardiac, and renal defects. Cancer risk was not increased with many common birth defects, including hypospadias, cleft lip with or without cleft palate, or hydrocephalus.

Conclusion

Children with some structural, non-chromosomal birth defects, but not others, have a moderately increased risk for childhood cancer. Information on such selective risk can promote more effective clinical evaluation, counseling, and research.     

Etiology of Diarrhea in Older Children,Adolescents and Adults: A Systematic Review

Background

Diarrhea is an important cause of morbidity and mortality in all regions of the world and among all ages, yet little is known about the fraction of diarrhea episodes and deaths due to each pathogen.

Methodology/Principal Findings

We conducted a systematic literature review to identify all papers reporting the proportion of diarrhea episodes with positive laboratory tests for at least one pathogen in inpatient, outpatient and community settings that met our inclusion and exclusion criteria. We identified a total of 25,701 papers with possible etiology data and after final screening included 22 papers that met all inclusion and exclusion criteria. Enterotoxigenic Escherichia coli and V. cholerae O1/O139 were the leading causes of hospitalizations. In outpatient settings, Salmonella spp., Shigella spp., and E. histolytica were the most frequently isolated pathogens.

Conclusions/Significance

This is the first systematic review which has considered the relative importance of multiple diarrhea pathogens. The few studies identified suggest that there is a great need for additional prospective studies around the world in these age groups to better understand the burden of disease and the variation by region.     

Tibialis Anterior Muscle Needle Biopsy and Sensitive Biomolecular Methods: A Useful Tool in Myotonic Dystrophy Type 1

Myotonic dystrophy type 1 (DM1) is a neuromuscular disorder caused by a CTG repeat expansion in 3’UTR of DMPK gene. This mutation causes accumulation of toxic RNA in nuclear foci leading to splicing misregulation of specific genes. In view of future clinical trials with antisense oligonucleotides in DM1 patients, it is important to set up sensitive and minimally-invasive tools to monitor the efficacy of treatments on skeletal muscle. A tibialis anterior (TA) muscle sample of about 60 mg was obtained from 5 DM1 patients and 5 healthy subjects through a needle biopsy. A fragment of about 40 mg was used for histological examination and a fragment of about 20 mg was used for biomolecular analysis. The TA fragments obtained with the minimally-invasive needle biopsy technique is enough to perform all the histopathological and biomolecular evaluations useful to monitor a clinical trial on DM1 patients.Key words: Myotonic dystrophy type 1, tibialis anterior; needle biopsy, alternative splicing, clinical trial     

Optical Saturation as a Versatile Tool to Enhance Resolution in Confocal Microscopy

One of the most actively developing areas in fluorescence microscopy is the achievement of spatial resolution below Abbe's diffraction limit, which restricts the resolution to several hundreds of nanometers. Most of the approaches in use at this time require a complex optical setup, a difficult mathematical treatment, or usage of dyes with special photophysical properties. In this work, we present a new, to our knowledge, approach in confocal microscopy that enhances the resolution moderately but is both technically and computationally simple. As it is based on the saturation of the transition from the ground state to the first excited state, it is universally applicable with respect to the dye used. The idea of the method presented is based on a principle similar to that underlying saturation excitation microscopy, but instead of applying harmonically modulated excitation light, the fluorophores are excited by picosecond laser pulses at different intensities, resulting in different levels of saturation. We show that the method can be easily combined with the concept of triplet relaxation, which by tuning the dark periods between pulses helps to suppress the formation of a photolabile triplet state and effectively reduces photobleaching. We demonstrate our approach imaging GFP-labeled protein patches within the plasma membrane of yeast cells.