Enzymhistochemical Localization of Alkaline Phosphatases in Germinal Cysts of the Testis of the Frog Rana temporaria (Anura)

Alkaline phosphatases have been demonstrated enzymhistochemically in the testis of the common frog Rana temporaria, caught in the month of January. The follicle cells in the cyst walls and the lamina propria of the seminiferous tubules showed strong enzymatic activity; also a weak activity was observed in the peripheral region of some of the germinal cells lying within the cysts. The possible interaction of alkaline phosphatases in the transport processes across the cyst walls has been discussed.     

Heterologous expression, purification, and kinetic comparison of the cytoplasmic and mitochondrial glyoxalase II enzymes, Glo2p and Glo4p, from Saccharomyces cerevisiae

The aims of the present study are (i) to purify a mitochondrial glyoxalase II to homogeneity for the first time from any organism and (ii) to compare its kinetic properties with those of the cytoplasmic enzyme. Both the cytoplasmic and the mitochondrial glyoxalases II from Saccharomyces cerevisiae, which are the products of two distinct genes, GLO2 and GLO4, were purified from yeast and in recombinant form from Escherichia coli. To obtain a higher protein yield (compared to wild-type expression) in yeast, the genes were placed under the control of the strong GAL1 promoter on a multicopy plasmid. Amino-terminal sequencing and molecular mass determination by MALDI-TOF mass spectrometry of the mitochondrial Glo4 protein revealed Met-11 of the primary translation product of the gene as the N-terminal amino acid. Judged by enzyme kinetic properties the recombinant and natural proteins were equivalent. The cytoplasmic and the mitochondrial enzyme differed in the pH dependence of the kinetic parameters for the main substrate, S-d-lactoylglutathione. Whereas the cytoplasmic protein showed a pronounced peak of enzyme activity between pH 7-8 and a continuous up to fivefold increase of the K(M) value with increasing pH (from 5. 5-9.0), the mitochondrial protein had a nearly constant K(M) value and an activity maximum over a broad pH range (6.5-9.0). The kinetic parameters (at pH 7.5) of both the cytoplasmic and the mitochondrial enzyme for S-D-lactoylglutathione were of the same order of magnitude as reported recently for the human and Arabidopsis thaliana enzymes which are presumably of cytoplasmic origin. However, both yeast enzymes showed a severalfold lower preference for the more hydrophobic substrate, S-d-mandeloylglutathione.     

Elevation of neutrophil-derived factors in patients after multiple trauma

Trauma represents one of the leading causes of death worldwide. Traumatic injuries elicit a dynamic inflammatory response with systemic release of inflammatory cytokines. Disbalance of this response can lead to systemic inflammatory response syndrome or compensatory anti-inflammatory response syndrome. As neutrophils play a major role in innate immune defence and are crucial in the injury-induced immunological response, we aimed to investigate systemic neutrophil-derived immunomodulators in trauma patients. Therefore, serum levels of neutrophil elastase (NE), myeloperoxidase (MPO) and citrullinated histone H3 (CitH3) were quantified in patients with injury severity scores above 15. Additionally, leukocyte, platelet, fibrinogen and CRP levels were assessed. Lastly, we analysed the association of neutrophil-derived factors with clinical severity scoring systems. Although the release of MPO, NE and CitH3 was not predictive of mortality, we found a remarkable increase in MPO and NE in trauma patients as compared with healthy controls. We also found significantly increased levels of MPO and NE on Days 1 and 5 after initial trauma in critically injured patients. Taken together, our data suggest a role for neutrophil activation in trauma. Targeting exacerbated neutrophil activation might represent a new therapeutic option for critically injured patients.     

Genetically engineered neural stem cells (NSCs) therapy for neurological diseases; state-of-the-art

Neural stem cells (NSCs) are multipotent stem cells with remarkable self-renewal potential and also unique competencies to differentiate into neurons, astrocytes, and oligodendrocytes (ODCs) and improve the cellular microenvironment. In addition, NSCs secret diversity of mediators, including neurotrophic factors (e.g., BDNF, NGF, GDNF, CNTF, and NT-3), pro-angiogenic mediators (e.g., FGF-2 and VEGF), and anti-inflammatory biomolecules. Thereby, NSCs transplantation has become a reasonable and effective treatment for various neurodegenerative disorders by their capacity to induce neurogenesis and vasculogenesis and dampen neuroinflammation and oxidative stress. Nonetheless, various drawbacks such as lower migration and survival and less differential capacity to a particular cell lineage concerning the disease pathogenesis hinder their application. Thus, genetic engineering of NSCs before transplantation is recently regarded as an innovative strategy to bypass these hurdles. Indeed, genetically modified NSCs could bring about more favored therapeutic influences post-transplantation in vivo, making them an excellent option for neurological disease therapy. This review for the first time offers a comprehensive review of the therapeutic capability of genetically modified NSCs rather than naïve NSCs in neurological disease beyond brain tumors and sheds light on the recent progress and prospect in this context.     

Characterisation of a repetitive DNA family from Entamoeba histolytica containing Saccharomyces cerevisiae ARS consensus sequences

Several repetitive DNA families were identified in Entamoeba histolytica DNA digested with Sau3AI. Characterisation of one of these repetitive DNA families showed the presence of multiple copies of Saccharomyces cerevisiae autonomously replicating sequence (ARS) core consensus sequences. The E. histolytica ARS consensus sequences allowed a yeast-integrating plasmid, YIP5, to replicate autonomously in S. cerevisiae. A 'bent DNA' fragment was located in one member of this E. histolytica repetitive DNA family.     

In vitro effect of mammalian pituitary hormones on germinal vesicle breakdown in oocytes of major carps, Labeo rohita, Cirrhinus mrigala, Catla catla and Cyprinus carpio

Among all the mammalian pituitary hormones, luteinizing hormone (LH) was the most potent in vitro inducer of oocyte maturation in L. rohita, C. mrigala, C. catla and C. carpio. It induced significant germinal vesicle breakdown (GVBD) at concentrations of 10, 1, 0.1 and 0.01 micrograms/ml. At the highest concentration used, LH induced 77.9 +/- 5.9, 73.8 +/- 4.6, 50.3 +/- 2.8 and 40.8 +/- 1.4% GVBD in oocytes of L. rohita, C. mrigala, C. catla and C. carpio, respectively. Among other hormones, follicle stimulating hormone (FSH) induced only a marginally significant GVBD (13.2 +/- 0.8%) in the oocytes of C. carpio, but not in other three species. Thyroid stimulating hormone (TSH), growth hormone (GH) and prolactin (PRL) had no effect on GVBD.     

Nonviral vector gene modification of stem cells for myocardial repair

Therapeutic angiogenesis and myogenesis restore perfusion of ischemic myocardium and improve left ventricular contractility. These therapeutic modalities must be considered as complementary rather than competing to exploit their advantages for optimal beneficial effects. The resistant nature of cardiomyocytes to gene transfection can be overcome by ex vivo delivery of therapeutic genes to the heart using genetically modified stem cells. This review article gives an overview of different vectors and delivery systems in general used for therapeutic gene delivery to the heart and provides a critical appreciation of the ex vivo gene delivery approach using genetically modified stem cells to achieve angiomyogenesis for the treatment of infarcted heart.     

Possibility of the Nonenzymatic Browning (Maillard) Reaction in the ISM

The possibility of the occurrence of the nonenzymatic browning reaction in the gaseous phase in the interstellar medium has been investigated by using Density Functional Theory computations. Mechanisms for the reactions between formaldehyde (Fald) + glycine (Gly), Fald + NH ₃ and Fald + methylamine (MeAm) have been proposed, and the possibility of the formation of different compounds in the proposed mechanisms has been evaluated through calculating the Gibb's free energy changes for different steps of the reaction, by following the total mass balance. The Fald + Gly reaction under basic conditions is found as the most favorable for producing 1-methyl-amino methene or 1-methyl-amino methelene (MAM). The reaction under acidic conditions is found to be the least favorable for producing MAM. The Fald + NH ₃ reaction is found to be plausible for the production of MeAm, which can participate by reaction with Fald, resulting in the formation of MAM.     

Mutation in the beta3 subunit of Guanine nucleotide-binding protein (GNB3) gene is not associated with Type II diabetes mellitus risk: a case–control study of a North Indian population

Context: Genetics play a major role in development and pathophysiology of Type 2 diabetes mellitus (T2DM).

Objective: To asses the association of Guanine nucleotide-binding protein (GNB3) (C825T) gene's polymorphism with T2DM.

Materials and methods: A case–control study including 400 North Indians was performed using Polymerase Chain Reaction–Restriction Fragment Length Polymorphism (PCR-RFLP) approach to analyze genetic polymorphism.

Results: No significant difference was observed in genotype and allele frequencies of GNB3 gene on comparing cases with controls.

Discussion: Our study is in agreement with studies on Polish, Japanese, Hispanic-American and Danish populations who observed no significant association between GNB3 (C825T) polymorphism and T2DM.

Conclusion: GNB3 (C825T) polymorphism is not associated with T2DM.     

Toward a methodical framework for comprehensively assessing forest multifunctionality

Biodiversity–ecosystem functioning (BEF) research has extended its scope from communities that are short‐lived or reshape their structure annually to structurally complex forest ecosystems. The establishment of tree diversity experiments poses specific methodological challenges for assessing the multiple functions provided by forest ecosystems. In particular, methodological inconsistencies and nonstandardized protocols impede the analysis of multifunctionality within, and comparability across the increasing number of tree diversity experiments. By providing an overview on key methods currently applied in one of the largest forest biodiversity experiments, we show how methods differing in scale and simplicity can be combined to retrieve consistent data allowing novel insights into forest ecosystem functioning. Furthermore, we discuss and develop recommendations for the integration and transferability of diverse methodical approaches to present and future forest biodiversity experiments. We identified four principles that should guide basic decisions concerning method selection for tree diversity experiments and forest BEF research: (1) method selection should be directed toward maximizing data density to increase the number of measured variables in each plot. (2) Methods should cover all relevant scales of the experiment to consider scale dependencies of biodiversity effects. (3) The same variable should be evaluated with the same method across space and time for adequate larger‐scale and longer‐time data analysis and to reduce errors due to changing measurement protocols. (4) Standardized, practical and rapid methods for assessing biodiversity and ecosystem functions should be promoted to increase comparability among forest BEF experiments. We demonstrate that currently available methods provide us with a sophisticated toolbox to improve a synergistic understanding of forest multifunctionality. However, these methods require further adjustment to the specific requirements of structurally complex and long‐lived forest ecosystems. By applying methods connecting relevant scales, trophic levels, and above‐ and belowground ecosystem compartments, knowledge gain from large tree diversity experiments can be optimized.