Gonadotropin-inhibitory hormone (GnIH) receptor gene is expressed in the chicken ovary: potential role of GnIH in follicular maturation

Gonadotropin-inhibitory hormone (GnIH), an RFamide peptide, has been found to inhibit pituitary LH secretion in avian and mammalian species. The gene encoding a putative receptor for GnIH (GnIHR) was recently identified in the chicken and Japanese quail brain and pituitary gland. GnIHR appears to be a seven-transmembrane protein belonging to a family of G-protein-coupled receptors. In the present study, we have characterized the expression of GnIHR mRNA in the chicken ovary and demonstrate that GnIHR may exert an inhibitory effect on ovarian follicular development. By RT-PCR, we detected GnIHR mRNA in the chicken testis and in the ovary, specifically both thecal and granulosa cell layers. Real-time quantitative PCR analysis revealed greater GnIHR mRNA quantity in theca cells of prehierarchial follicles compared with that of preovulatory follicles. GnIHR mRNA quantity was significantly decreased in sexually mature chicken ovaries versus ovaries of sexually immature chickens. Estradiol (E(2)) and/or progesterone (P(4)) treatment of sexually immature chickens significantly decreased ovarian GnIHR mRNA abundance. Treatment of prehierarchial follicular granulosa cells in vitro with chicken GnIH peptide significantly decreased basal but not FSH-stimulated cellular viability. Collectively, our results indicate that the ovarian GnIHR is likely to be involved in ovarian follicular development. A decrease in ovarian GnIHR mRNA abundance due to sexual maturation or by E(2) and/or P(4) treatment would implicate an inhibitory role for GnIHR in ovarian follicular development. Furthermore, GnIH may affect follicular maturation by decreasing the viability of prehierarchial follicular granulosa cells through binding to GnIHR.     

Investigation of the role of tyrosine-114 in the activity of human O6-alkylguanine-DNA alkyltranferase

Tyrosine-114 is one of 13 totally conserved amino acids in all known sequences of O6-alkylguanine-DNA alkyltransferase (AGT). The importance of this amino acid in repair of alkylated DNA by AGT was studied by changing it to phenylalanine (F), alanine (A), threonine (T), or glutamic acid (E) in human AGT. The activities of the mutant proteins were then compared to those of the wild type with regard to abilities to do the following: (a) protect Escherichia coli from the methylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG); (b) repair methylated DNA in vitro; (c) bind to oligodeoxynucleotides containing O6-methylguanine; and (d) react with the low molecular weight pseudosubstrate, O6-benzylguanine. When expressed at high levels in E. coli strain GWR109, lacking endogenous AGT, the wild type and the Y114F mutant were highly effective in reducing mutations and cell killing by MNNG. The Y114A mutant had a much smaller protective effect, and mutants Y114T and Y114E were inactive. Purified preparations of all four AGT mutants showed an approximately similar degree (74-120-fold) of reduction in the rate of reaction with O6-benzylguanine. In contrast, the degree of reduction in activity toward methylated DNA substrates in vitro varied according to the mutation with the more conservative Y114F producing only a 30-fold reduction and the most drastic change of Y114E abolishing activity completely. Alteration Y114A produced a 1000-fold reduction whereas Y114T reduced activity by 10000-fold. All of the mutations affected the binding of AGT to single- or double-stranded oligodeoxynucleotides containing O6-methylguanine. The extent of increase in the Kd varied according to the amino acid with 2-5-fold (F), 7-11-fold (A), 167-200-fold (T), and 600-1000-fold (E) increases. These results are consistent with tyrosine-114 playing a role both in the binding of AGT to its DNA substrate and in facilitating the transfer of the alkyl group. It is probable that AGT resembles other DNA repair proteins in bringing about a "flipping out" of the target base from the DNA helix. Tyrosine-114 is therefore an excellent candidate for a key role in the interaction with the flipped O6-methylguanine. The results also show that when large amounts of AGT are produced in the cell, substantial decreases in the efficiency with which AGT can repair methylated DNA do not prevent the ability to protect E. coli from toxic alkylating agents. Mutant Y114F, whose activity was reduced by 30-fold, was equal to wild-type AGT in bringing about this protection.     

Thermo-mechanical treatments of Sc- and Mg-modified Al–Cu alloy welds

High-strength heat-treatable aluminum alloy AA2219 finds application in aerospace industries. Though it has good weldability, with alternating current–tungsten inert gas welding, the joint efficiency obtained is only 40%, particularly in thicker plates. In the present study, an attempt has been made to improve the weld metal properties by modifying the chemistry of fusion zone and post-weld thermo-mechanical treatments. Fillers were made through casting route by melting conventional 2319 filler with Sc and Mg. Two levels of Sc (0.3% and 0.6%) and four levels of Mg (0.3% to 0.6%) were varied. Compressive deformation was done on the fusion zone of the weld to get three levels of percentage of reduction (4%, 8%, and 12%). As welded specimens and welds after compressive deformation, those were subjected to post-weld aging treatments at 190ºC for different periods up to 100 h. Compressive deformation on the welds made with modified filler of 2319 with Sc and Mg resulted in significant improvement in the weld metal strength.     

Antifungal activity of Azotobacter nigricans against trichothecene-producing Fusarium species associated with cereals

Food Science and Biotechnology - Antifungal efficacy of Azotobacter nigricans on Fusarium infection, total seedlings mass, root and shoot length, and seed germination in maize, sorghum, and wheat...     

Enhancing total agility level through assessment and product mapping: A case study in the manufacturing of refrigeration air dryer

The world’s manufacturing community has been questing for ways to face the onslaught of competition. One of those ways is the adoption of agile manufacturing (AM) paradigm. AM paradigm enables a company to quickly respond to the customers’ dynamic demands. In order to implement AM paradigm, a model named “model for enhancing total agility level” (METAL) is proposed in this paper. METAL enunciates the assessing of the total agility level (TAL), identifying the weak AM criteria and subsequently strengthening them. The practicality of METAL has been explored in an air dryer manufacturing company. During this case study, refrigeration air dryer was considered as AM capable product. After assessment, three weak AM criteria were identified. Proposals were drawn to strengthen these three weak AM criteria. These proposals envisage the strengthening of the weak AM criteria through the removal of nonvalue adding activities, utilization of mathematical models, and creation of web portal. The reassessment has indicated the possibility of enhancing the TAL value in the above company through the implementation of these proposals. The experience of carrying out this research has revealed that the deployment of METAL would facilitate the contemporary companies to systematically infuse AM paradigm and enhance their TAL values.     

Attribute-based key-insulated signature for boolean formula

In order to minimize the impact of secret signing key exposure in attribute-based signature scenario, we construct an attribute-based key-insulated signature (ABKIS) scheme for expressive monotone boolean function access structures utilizing only four pairing operations in verification process and making the signature length constant, that is, the number of pairings required for signature verification and the size of signature are independent of the size of attribute set participated in the respective process. The (strong) key-insulated selective security of our ABKIS scheme is reduced to the computational Diffie–Hellman Exponent problem without using any random oracles. The proposed construction attains signer privacy, which is a fundamental requirement of the signature schemes in the attribute-based setting.     

Reaction of O6-benzylguanine-resistant mutants of human O6-alkylguanine-DNA alkyltransferase with O6-benzylguanine in oligodeoxyribonucleotides

Inactivation of the human DNA repair protein, O6-alkylguanine-DNA alkyltransferase (AGT), by O6-benzylguanine renders tumor cells susceptible to killing by alkylating agents. AGT mutants resistant to O6-benzylguanine can be made by converting Pro140 to an alanine (P140A) or Gly156 to an alanine (G156A). These mutations had a much smaller effect on the reaction with O6-benzylguanine when it was incorporated into a short single-stranded oligodeoxyribonucleotide. Such oligodeoxyribonucleotides could form the basis for the design of improved AGT inhibitors. AGT and mutants P140A and G156A preferentially reacted with O6-benzylguanine when incubated with a mixture of two 16-mer oligodeoxyribonucleotides, one containing O6-benzylguanine and the other, O6-methylguanine. When the 6 amino acids located in positions 159-164 in AGT were replaced by the equivalent sequence from the Escherichia coli Ada-C protein (mutant AGT/6ada) the preference for benzyl repair was eliminated. Further mutation incorporating the P140A change into AGT/6ada giving mutant P140A/6ada led to a protein that resembled Ada-C in preference for the repair of methyl groups, but P140A/6ada did not differ from P140A in reaction with the free base O6-benzylguanine. Changes in the AGT active site pocket can therefore affect the preference for repair of O6-benzyl or -methyl groups when present in an oligodeoxyribonucleotide without altering the reaction with free O6-benzylguanine.     

Improvement of phone recognition accuracy using speech mode classification

In this work, we have developed a speech mode classification model for improving the performance of phone recognition system (PRS). In this paper, we have explored vocal tract system, excitation source and prosodic features for development of speech mode classification (SMC) model. These features are extracted from voiced regions of a speech signal. In this study, conversation, extempore, and read speech are considered as three different modes of speech. The vocal tract component of speech is extracted using Mel-frequency cepstral coefficients (MFCCs). The excitation source features are captured through Mel power differences of spectrum in sub-bands (MPDSS) and residual Mel-frequency cepstral coefficients (RMFCCs) of the speech signal. The prosody information is extracted from pitch and intensity. Speech mode classification models are developed using above described features independently, and in fusion. The experiments carried out on Bengali speech corpus to analyze the accuracy of the speech mode classification model using the artificial neural network (ANN), naive Bayes, support vector machines (SVMs) and k-nearest neighbor (KNN). We proposed four classification models which are combined using maximum voting approach for optimal performance. From the results, it is observed that speech mode classification model developed using the fusion of vocal tract system, excitation source and prosodic features of speech, yields the best performance of 98%. Finally, the proposed speech mode classifier is integrated to the PRS, and the accuracy of phone recognition system is observed to be improved by 11.08%.