Mitochondrial dysfunction and oxidative stress caused by cryopreservation in reproductive cells

Mitochondria, fundamental organelles in cell metabolism, and ATP synthesis are respon-sible for generating reactive oxygen species (ROS), calcium homeostasis, and cell death. Mitochon-dria produce most ROS, and when levels exceed the antioxidant defenses, oxidative stress (OS) is generated. These changes may eventually impair the electron transport chain, resulting in decreased ATP synthesis, increased ROS production, altered mitochondrial membrane permeability, and dis-ruption of calcium homeostasis. Mitochondria play a key role in the gamete competence to facilitate normal embryo development. However, iatrogenic factors in assisted reproductive technologies (ART) may affect their functional competence, leading to an abnormal reproductive outcome. Cry-opreservation, a fundamental technology in ART, may compromise mitochondrial function leading to elevated intracellular OS that decreases sperm and oocytes’ competence and the dynamics of fertilization and embryo development. This article aims to review the role played by mitochondria and ROS in sperm and oocyte function and the close, biunivocal relationships between mitochon-drial damage and ROS generation during cryopreservation of gametes and gonadal tissues in different species. Based on current literature, we propose tentative hypothesis of mechanisms involved in cryopreservation-associated mitochondrial dysfunction in gametes, and discuss the role played by antioxidants and other agents to retain the competence of cryopreserved reproductive cells and tissues

Seedling selection in open pollinated genotypes of cashew (Anacardium occidentale)

In a highly heterozygous crop like cashew (Anacardium occidentale L.), selection of high yielding trees from a seedling population of existing varieties is one of the ways of development of variety. Hence an experiment was undertaken to study the extent of variability in seedlings of six popular genotypes, viz. NRCC Sel 2, Vengurle 4, Vridhachalam 3, Bhaskara, VTH 174 and VTH 30/4 and to identify promising trees. Eighty open pollinated seeds each from trees belonging to six genotypes were collected; seedlings were raised and planted during 2007 at Directorate of Cashew Research, Experimental Station, Shantigodu. Growth parameters like trunk girth, tree height, tree spread and nut yield per tree were recorded. The progeny performance in different varieties revealed that the mean nut yield per tree was highest with 2.46 kg in Vridhachalam 3. With regard to individual tree performance tree number 480 belonging to VTH 30/4 recorded highest yield of 6.9 kg nuts/tree. The frequency distribution patterns showed that data of trunk girth was moderately negatively skewed implying increasing alleles are in slight excess and dominant for this trait. Whereas tree height and tree spread distributions were moderately positively skewed indicating decreasing alleles are in slight excess and dominant for these traits. The nut yield showed highly positively skewed distribution revealing decreasing alleles are in excess and dominant. The kurtosis was very high for nut yield indicating that yield variability is due to a few extreme differences from the mean. It was observed that trunk girth and tree spread were positively correlated with nut yield. The study could identify one promising seedling (T No.480), a seedling progeny of VTH- 30/4 in terms of nut yield

Angle-Resolved Environmental X-Ray Photoelectron Spectroscopy: A New Laboratory Setup for Photoemission Studies at Pressures up to 0.4 Torr

The paper presents the development and demonstrates the capabilities of a new laboratory-based environmental X-ray photoelectron spectroscopy system incorporating an electrostatic lens and able to acquire spectra up to 0.4 Torr. The incorporation of a two-dimensional detector provides imaging capabilities and allows the acquisition of angle-resolved data in parallel mode over an angular range of 14° without tilting the sample. The sensitivity and energy resolution of the spectrometer have been investigated by analyzing a standard Ag foil both under high vacuum (10−8 Torr) conditions and at elevated pressures of N2 (0.4 Torr). The possibility of acquiring angle-resolved data at different pressures has been demonstrated by analyzing a silicon/silicon dioxide (Si/SiO2) sample. The collected angle-resolved spectra could be effectively used for the determination of the thickness of the native silicon oxide layer

Effect of growth regulators and micronutrients on quality parameters in cashew (Anacardium occidentale L.)

Cashew (Anacardium occidentale L.) is an important tropical nut crop of social and economic importance worldwide. However, the crop is threatened with the low yield. In the present study, an attempt was made to test the effects of plant growth hormones as well as micronutrients on nut and apple quality of cashew var. Bhaskara. Significant differences in kernel weight, shelling percentage, carbohydrates and starch content of cashew kernel and juice content of cashew apple were observed with the foliar application of growth hormones and micronutrients. The foliar application of ethrel @ 50 ppm increased shelling percentage (35.8%), carbohydrate content (21.63%), sugar content (6.26%), protein content (32.4%), starch content (31.42%), juice content (78.3%) and total soluble solids (120 Brix). Further, the foliar spray of zinc sulphate (0.5%) + borax (0.1%) increased shelling (36.13%), protein content (32.15%), starch content (32.03%) among all the treatments tested. Furthermore, higher cashew apple juice content (78%) and total soluble solids (120Brix) was also recorded with the foliar spray of zinc sulphate (0.5%) + borax (0.1%)

When the optimal is not the best: parameter estimation in complex biological models

Background: The vast computational resources that became available during the past decade enabled the development and simulation of increasingly complex mathematical models of cancer growth. These models typically involve many free parameters whose determination is a substantial obstacle to model development. Direct measurement of biochemical parameters in vivo is often difficult and sometimes impracticable, while fitting them under data-poor conditions may result in biologically implausible values. Results: We discuss different methodological approaches to estimate parameters in complex biological models. We make use of the high computational power of the Blue Gene technology to perform an extensive study of the parameter space in a model of avascular tumor growth. We explicitly show that the landscape of the cost function used to optimize the model to the data has a very rugged surface in parameter space. This cost function has many local minima with unrealistic solutions, including the global minimum corresponding to the best fit. Conclusions: The case studied in this paper shows one example in which model parameters that optimally fit the data are not necessarily the best ones from a biological point of view. To avoid force-fitting a model to a dataset, we propose that the best model parameters should be found by choosing, among suboptimal parameters, those that match criteria other than the ones used to fit the model. We also conclude that the model, data and optimization approach form a new complex system, and point to the need of a theory that addresses this problem more generally

Limits of phytosanitation and host plant resistance towards the control of cassava viruses in Uganda

Published online: 30 Sept 2017Cassava (Manihot esculenta Crantz) and the viruses that infect it, notably cassava mosaic virus and cassava brown streak viruses, have a unique history of co-evolution and co-existence. While cassava originated in South America, both viruses and the diseases they cause have largely been limited to the East African region, where they have, and continue to be key yield-robbing stresses. For sustainable control, we assume that deployment of resistant varieties when carefully combined with phytosanitation will combat these viruses. We have thus generated empirical data and tested the limits, i.e., how long this strategy can last. This entailed the comparison of elite cassava varieties, one set of virus-indexed tissue culture plantlets, and the other set, re-cycled planting materials under farmer’s cyclic propagation for 6-23 years. Trials were established at diverse sites in Uganda. We observed that both officially-released and unofficially-released cassava varieties are common in farmer’s fields; these varieties have varying susceptibility levels to viruses. Worrisome was that some officially-released varieties like NASE 3 registered cassava mosaic disease (CMD) incidences of up to 71% (virus-indexed), which was not any different from its re-cycled counterparts. Other varieties like NASE 14 have maintained high levels of CMD resistance six years after official release. Predominant re-cycled cassava varieties notably TME 204, I92/0057, TME 14, and to a limited extent NASE 14, are key reservoirs for cassava brown streak disease (CBSD) associated viruses. These findings highlight the limits of phytosanitation, i.e., in areas like Kaberamaido associated with high CMD pressure, varieties NASE 1 and NASE 3 can not be recommended; on the contrary, these varieties can be deployed in Kalangala, where they can survive with phytosanitation. And for CBSD, the findings justify the urgent need for phytosanitation (community-led) and development of varieties with higher levels of resistance and/or tolerance, as no immune variety has so far been identified

Switching Transport through Nanopores with pH-Responsive Polymer Brushes for Controlled Ion Permeability

Several nanoporous platforms were functionalized with pH-responsive poly(methacrylic acid) (PMAA) brushes using surface-initiated atom transfer radical polymerization (SI-ATRP). The growth of the PMAA brush and its pH-responsive behavior from the nanoporous platforms were confirmed by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and atomic force microscopy (AFM). The swelling behavior of the pH-responsive PMAA brushes grafted only from the nanopore walls was investigated by AFM in aqueous liquid environment with pH values of 4 and 8. AFM images displayed open nanopores at pH 4 and closed ones at pH 8, which rationalizes their use as gating platforms. Ion conductivity across the nanopores was investigated with current–voltage measurements at various pH values. Enhanced higher resistance across the nanopores was observed in a neutral polymer brush state (lower pH values) and lower resistance when the brush was charged (higher pH values). By adding a fluorescent dye in an environment of pH 4 or pH 8 at one side of the PMAA-brush functionalized nanopore array chips, diffusion across the nanopores was followed. These experiments displayed faster diffusion rates of the fluorescent molecules at pH 4 (PMAA neutral state, open pores) and slower diffusion at pH 8 (PMAA charged state, closed pores) showing the potential of this technology toward nanoscale valve applications

DeadEasy Mito-Glia: Automatic Counting of Mitotic Cells and Glial Cells in Drosophila

Cell number changes during normal development, and in disease (e.g., neurodegeneration, cancer). Many genes affect cell number, thus functional genetic analysis frequently requires analysis of cell number alterations upon loss of function mutations or in gain of function experiments. Drosophila is a most powerful model organism to investigate the function of genes involved in development or disease in vivo. Image processing and pattern recognition techniques can be used to extract information from microscopy images to quantify automatically distinct cellular features, but these methods are still not very extended in this model organism. Thus cellular quantification is often carried out manually, which is laborious, tedious, error prone or humanly unfeasible. Here, we present DeadEasy Mito-Glia, an image processing method to count automatically the number of mitotic cells labelled with anti-phospho-histone H3 and of glial cells labelled with anti-Repo in Drosophila embryos. This programme belongs to the DeadEasy suite of which we have previously developed versions to count apoptotic cells and neuronal nuclei. Having separate programmes is paramount for accuracy. DeadEasy Mito-Glia is very easy to use, fast, objective and very accurate when counting dividing cells and glial cells labelled with a nuclear marker. Although this method has been validated for Drosophila embryos, we provide an interactive window for biologists to easily extend its application to other nuclear markers and other sample types. DeadEasy MitoGlia is freely available as an ImageJ plug-in, it increases the repertoire of tools for in vivo genetic analysis, and it will be of interest to a broad community of developmental, cancer and neuro-biologists