Совершенствование управления персоналом с использованием программных систем на примере Нефтеюганского филиала АО «ССК»

Объектом исследования является: программная система управления персоналом организации Нефтеюганского филиала АО "Сибирская Сервисная Компания". Цель работы – выявление особенностей системы программного обеспечения в управление персоналом в НФ АО "ССК" и разработка мероприятий по повышению эффективности работы с персоналом с использованием нового программного обеспечения. В результате исследования:предложена программная система "Босс-кадровик" для совершенствования процесса управления персоналом на анализируемом предприятии. Актуальность темы состоит в оптимизации процесса управления персоналом, за счет внедрения программной системы для исследуемого предприятия Нефтеюганского филиала АО "ССК".The subject of the study is: a software system for personnel management of the Nefteyugansk branch of JSC Siberian Service Company. The purpose of the work is to identify the features of the software system in personnel management in the NF of "SSK" and to develop measures to improve the efficiency of work with personnel using new software. As a result of the research: the "Boss-kadrovik" software system was proposed to improve the process of personnel management at the analyzed enterprise. The topicality of the topic is to optimize the process of personnel management, through the introduction of a software system for the company under study in the Nefteyugansk branch of SSK

Reactive Oxygen Species Facilitate Translocation of Hormone Sensitive Lipase to the Lipid Droplet During Lipolysis in Human Differentiated Adipocytes

In obesity, there is an increase in reactive oxygen species (ROS) within adipose tissue caused by increases in inflammation and overnutrition. Hormone sensitive lipase (HSL) is part of the canonical lipolytic pathway and critical for complete lipolysis. This study hypothesizes that ROS is a signal that integrates regulation of lipolysis by targeting HSL. Experiments were performed with human differentiated adipocytes from the subcutaneous depot. Antioxidants were employed as a tool to decrease ROS, and it was found that scavenging ROS with diphenyliodonium, N-acetyl cysteine, or resveratrol decreased lipolysis in adipocytes. HSL phosphorylation of a key serine residue, Ser552, as well as translocation of this enzyme from the cytosol to the lipid droplet upon lipolytic stimulation were both abrogated by scavenging ROS. The phosphorylation status of other serine residues on HSL were not affected. These findings are significant because they document that ROS contributes to the physiological regulation of lipolysis via an effect on translocation. Such regulation could be useful in developing new obesity therapies

Κλωνοποίηση και μελέτη της λειτουργίας του γονιδίου της ορμονοευαίσθητης λιπάσης (HSL) του προβάτου

Hormone Sensitive Lipase (HSL) is a highly regulated enzyme that mediates lipolysis in adipocytes. HSL enzymatic activity is increased by adrenergic agonists, such as catecholamines and glucagons, which induce cyclic AMP (cAMP) intracellular production, subsequently followed by the activation ofProtein Kinase A (PKA) and its downstream signalling cascade reactions. Since HSL constitutes the key enzyme in the regulation of lipid stores and the only enzyme being subjected to hormonal regulation [in terms of the recently identified Adipose Triglyceride Lipase (ATGL)], the ovine Hormone Sensitive Lipase (ovHSL) full-length cDNA clones were isolated, using a Polymerase Chain Reaction-based (PCR) strategy. The two isolated isoforms ovHSL-A and ovHSL-B contain two highly homologous Open Reading Frame (ORF) regions of 2089 bp and 2086 bp, respectively, the latter having been missed the 688th triplet coding for glutamine (ΔQ⁶⁸⁸). The putative 695 and 694 amino acid respective sequences bear strong homologies with other HSL protein family members. Southern blotting analysis revealed that HSL is represented as a single copy gene in the ovine genome, while Reverse Transcription-PCR (RT-PCR) approaches unambiguously dictated its variable transcriptional expression profile in the different tissues examined. Interestingly, as undoubtedly corroborated by both RTPCR and Western blotting analysis, ovHSL gene expression is notably enhanced in the adipose tissue during the fasting period, when lipolysis is highly increased in ruminant species. Based on the crystal structure of an Archaeoglobus fulgidus enzyme, a three-dimensional (3D) molecular model of the ovHSL putative catalytic domain was constructed, thus providing an inchoative insight into understanding the enzymatic activity and functional regulation mechanisms of the ruminant HSL gene product(s). In order to acquire detailed knowledge with regard to the mechanisms regulating ovine HSL gene transcription activity, we initially isolated and cloned the 5' proximal and distal promoter regions through a genome walking approach, with the utilization of the previously characterized ovHSL cDNAs. As evinced by BLAST analysis and a multiple alignment procedure, the isolated genomic fragment of 2744 bp appeared to contain the already specified 5'-untranslated region (5'-UTR), which was surprisingly interrupted by a relatively large intron of 1448 bp. Regarding the upstream remaining part of 1224 bp, it was demonstrated to represent a TATA-less promoter area, harboring several cfs-regulatory elements that could be putatively recognized by relatively more general transcription factors, mainly including Stimulating protein 1 (Sp1), CCAAT box Binding Factors (CBFs), Activator Protein 2 (AP2) and Glucocorticoid Receptor (GR), as well as other cis-acting regions denominated as Insulin Response Element (IRE), Glucose Response Element (GRE), Fat Specific Element (FSE) and cAMP Response Element (CRE), which could likely function in a condition-/hormone-dependent fashion. When different genomic fragments were directionally (5' to 3') cloned into a suitable reporter vector upstream of a promoter-less luciferase gene and transiently transfected into 3T3-L1 (mouse fibroblasts) as well as T24 (human bladder cancer) cell lines, strong promoter activities were unambiguously detected, with the -140/+18 nucleotide sequence bearing the highest transcriptional response, thus indicating that the 1224 bp 5' flanking region, isolated by genome walking, veritably contains the ovHSL gene promoter. Of particular significance are the observations that the functional promoter fragments could trigger the transcriptional activity of luciferase gene only under high concentration of glucose conditions in both cell lines, whereas the respective insulin responses seemed to follow a cell type-specific pattern.Η ορμονοευαίσθητη λιπάση (Hormone Sensitive Lipase-HSL) διαδραματίζει σημαντικό ρόλο στη διαδικασία της λιπόλυσης στα αγροτικά ζώα (μηρυκαστικά, χοίρος), καθώς αποτελεί το ένζυμο-κλειδί, το οποίο βρίσκεται κάτω από ορμονικό έλεγχο, συμβάλλοντας καταλυτικά στη διάσπαση των τριγλυκεριδίων, της κύριας μορφής αποθήκευσης ενέργειας, προς διγλυκερίδια και απελευθέρωση λιπαρού οξέος. Η ορμονοευαίσθητη λιπάση ενεργοποιείται ύστερα από επίδραση στα κύτταρα του λιπώδους ιστού ορμονών, όπως η αδρεναλίνη και η γλυκαγόνη. Οι ορμόνες αυτές διεγείρουν κατάλληλους υποδοχείς, οι οποίοι ενεργοποιούν το ένζυμο αδενυλική κυκλάση (αδενυλοκυκλάση) της μεμβράνης των λιποκυττάρων. Η αυξημένη συγκέντρωση της κυκλικής μονοφωσφορικής αδενοσίνης (cAMP) στη συνέχεια, διεγείρει την πρωτεϊνική κινάση Α, η οποία ενεργοποιεί μέσω φωσφορυλίωσης, τη λιπάση. Στόχο της παρούσας διδακτορικής διατριβής αποτέλεσε η απομόνωση και ο μοριακός χαρακτηρισμός του γονιδίου που κωδικεύει την ορμονοευαίσθητη λιπάση στο πρόβατο. Η εφαρμογή της αλυσιδωτής αντίδρασης της πολυμεράσης οδήγησε στην απομόνωση του πλήρους μήκους cDNA που κωδικεύει την HSL. Συγκεκριμένα, απομονώθηκαν και κλωνοποιήθηκαν δύο κλώνοι-ισόμορφα ovHSL-A και ovHSL-B συνολικού μήκους 2248 bp και 2221 bp αντίστοιχα, που κωδικεύουν το συγκεκριμένο ένζυμο, διατηρώντας ανοιχτό αναγνωστικό πλαίσιο 695 και 694 αμινοξέων, αντίστοιχα. Οι διαφορές των δύο κλώνων έγκεινται στην ύπαρξη ενός επιπλέον τρινουκλεοτιδίου στο ισόμορφο ovHSL-A που αντιστοιχεί στο αμινοξύ γλουταμίνη (Q), καθώς και στην επιπρόσθετη περιοχή μήκους 24 bp την οποία φέρει το ίδιο ισόμορφο και αποτελεί τμήμα της μη μεταφραζόμενης περιοχής (3' UTR) του γονιδίου. Στοίχιση της αμινοξικής αλληλουχίας των δύο κλώνων με τις αντίστοιχες αλληλουχίες των πρωτεϊνών άλλων θηλαστικών (άνθρωπος, ποντικός, αρουραίος, χοίρος, αγελάδα), έδειξε υψηλό ποσοστό συντήρησης. Παράλληλα, η προσπάθεια εύρεσης του αριθμού των αντιγράφων του υπό μελέτη γονιδίου, φανέρωσε ότι το συγκεκριμένο γονίδιο απαντά ως μοναδιαίο αντίγραφο (single copy gene) εντός του γονιδιώματος του προβάτου. Επιπλέον, μελετήθηκε η έκφραση του γονιδίου της ορμονοευαίσθητης λιπάσης σε διάφορους ιστούς προβάτου και βρέθηκε ότι το γονίδιο εκφράζεται σε μεγάλο βαθμό τόσο σε επίπεδο mRNA όσο και σε επίπεδο πρωτεΐνης στο λιπώδη ιστό, σε συνθήκες περιορισμένης διατροφής, περίοδος κατά την οποία η διαδικασία της λιπόλυσης είναι ιδιαίτερα αυξημένη. Το γονίδιο παρουσίασε παρόμοιο, χαμηλό πρότυπο έκφρασης στην περίπτωση των επινεφριδίων και των όρχεων, ενώ σημαντικά μειωμένο ήταν το επίπεδο έκφρασης στην περίπτωση των υπόλοιπων ιστών, παρουσιάζοντας μικρές διαφοροποιήσεις. Επομένως, η κατάσταση του υπό εξέταση οργανισμού και η επίδραση εξωγενών παραγόντων, όπως η διατροφή, αποτελούν καθοριστικούς παράγοντες με καταλυτική επίδραση στην έκφραση του γονιδίου της ορμονοευαίσθητης λιπάσης. Η εύρεση χαρακτηριστικών δομών και λειτουργικών περιοχών της πρωτεΐνης του υπό μελέτη γονιδίου, σύμφωνα με τα ήδη προσδιορισμένα πρωτεϊνικά μόρια άλλων οργανισμών, βοήθησε στην κατασκευή ενός τρισδιάστατου δομικού πρωτεϊνικού μοντέλου της ορμονοευαίσθητης λιπάσης. Το μοντέλο αποτελεί ένα αρχικό πρότυπο για την περαιτέρω ανάλυση των δομικών περιοχών και τη μελέτη δομικών αλλαγών από μελλοντικές μεταλλάξεις που πιθανόν να εντοπιστούν στην περιοχή του ενεργού κέντρου με επίπτωση στη λειτουργική δράση του ενζύμου. Πέραν των παραπάνω, μελετήθηκε η μεταγραφική ρύθμιση και έκφραση της λειτουργικής περιοχής του υποκινητή του γονιδίου της HSL. Η απομόνωση του υποκινητή πραγματοποιήθηκε με την τεχνική του χρωμοσωμικού βαδίσματος, απομονώνοντας εν τέλει περιοχή 1224 bp. Η in silico ανάλυση της περιοχής αυτής, ανέδειξε την παρουσία αρκετών θέσεων πιθανής πρόσδεσης μεταγραφικών παραγόντων όπως είναι οι: Spi1(Stimulating protein 1), SRE (Sterol Regulatory Element), AP2 (Activator Protein 2), FSE (Fat Specific Element), GBP (G-Box Binding Proteins) CCAAT (cellular and viral CCAAT Box) ARE (Adipocyte Response Element), GR (Glucocorticoid Receptor), CRE (cAMP Response Element) IRE (Insulin Response Element), STRE (Stress Response Element), GRE (Glucose Response Element), SE (Steroidogenic Element), PRE (Preadipocyte Regulatory Element), GATA-1 (GATA Binding Factor 1), GATA-2 (GATA Binding Factor 2), PPRE (Peroxisome Proliferator-activated Receptor Element) και HSF (Heat Shock Factor), οι οποίοι εμπλέκονται στη μεταγραφική ρύθμιση του υπό μελέτη γονιδίου και κατ’ επέκταση στη ρύθμιση της διαδικασίας της λιπόλυσης. Η συγκριτική μελέτη των περιοχών του υποκινητή μεταξύ προβάτου και ανθρώπου, ως προς τους βασικούς μεταγραφικούς παράγοντες, κατέγραψε την παρουσία cis-λειτουργικών στοιχείων, αναδεικνύοντας ορισμένες διαφορές ως προς τον αριθμό και την ακριβή θέση που καταλαμβάνει ο εκάστοτε μεταγραφικός παράγοντας στην 5' ρυθμιστική περιοχή του γονιδίου. Οι διαφορές αυτές εξασφαλίζουν, ταυτόχρονα, μια πρώτη εικόνα για τη μεταγραφική ρύθμιση και λειτουργία του συγκεκριμένου γονιδίου μεταξύ μηρυκαστικών και μονογαστρικών ειδών. Παράλληλα, η μελέτη της μεταγραφικής απόκρισης συγκεκριμένων cis-ρυθμιστικών περιοχών και λειτουργικών στοιχείων του υποκινητή του γονιδίου, με την παροδική διαμόλυνση ευκαρυωτικών κυττάρων και την επίδραση διαφορετικών «παραγόντων-φορέων», αποτελεί μια πρώτη προσέγγιση για τη ρύθμιση της λειτουργικότητας της ευρύτερης περιοχής του υποκινητή, αποσαφηνίζοντας τα όρια της «ελάχιστης» δομικά cis-ρυθμιστικής περιοχής που απαιτείται για την ελεγχόμενη γονιδιακή δραστηριότητα και καθορίζοντας τους μεταγραφικούς παράγοντες με καταλυτικό ρόλο στη μεταγραφική ρύθμιση του γονιδίου. Τέλος, η μελέτη τυχών παρατηρούμενων μεταλλάξεων στην περιοχή του υποκινητή, οι οποίες είναι άμεσα συνυφασμένες με παραγωγικές ιδιότητες του ζώου, όπως η εναπόθεση λίπους, ασκώντας επίδραση στη μεταγραφική ρύθμιση του γονιδίου, θα αποτελούσε έναν ενδιαφέρον τομέα έρευνας, καθορίζοντας νέους βελτιωτικούς στόχους για τη ζωική παραγωγή

The resurgence of Hormone-Sensitive Lipase (HSL) in mammalian lipolysis

The ability to store energy in the form of energy-dense triacylglycerol and to mobilize these stores rapidly during periods of low carbohydrate availability or throughout the strong metabolic demand is a highly conserved process, absolutely essential for survival. In the industrialized world the regulation of this pathway is viewed as an important therapeutic target for disease prevention. Adipose tissue lipolysis is a catabolic process leading to the breakdown of triacylglycerols stored in fat cells, and release of fatty acids and glycerol. Mobilization of adipose tissue fat is mediated by the MGL, HSL and ATGL, similarly functioning enzymes. ATGL initiates lipolysis followed by the actions of HSL on diacylglycerol, and MGL on monoacylglycerol. HSL is regulated by reversible phosphorylation on five critical residues. Phosphorylation alone, however, is not enough to activate HSL. Probably, conformational alterations and a translocation from the cytoplasm to lipid droplets are also involved. In accordance, Perilipin functions as a master regulator of lipolysis, protecting or exposing the triacylglycerol core of a lipid droplet to lipases. The prototype processes of hormonal lipolytic control are the β-adrenergic stimulation and suppression by insulin, both of which affect cytoplasmic cyclic AMP levels. Lipolysis in adipocytes is an important process in the management of body energy reserves. Its deregulation may contribute to the symptoms of type 2 diabetes mellitus and other pathological situations. We, herein, discuss the metabolic regulation and function of lipases mediating mammalian lipolysis with a focus on HSL, quoting newly identified members of the lipolytic proteome. © 2011 Elsevier B.V

Soya protein hydrolysates modify the expression of various pro-inflammatory genes induced by fatty acids in ovine phagocytes

The objective of the present study was to test the hypothesis that fatty acids are the circulating mediators acting in a pro-inflammatory manner towards activated circulating ovine monocyte/macrophages and neutrophils. Furthermore, whether soya protein hydrolysates (SPH) inhibit the fatty acid-induced increase in the production of pro-inflammatory responses by ovine phagocytes was tested in vitro. All the fatty acids tested (myristic, palmitic, palmitoleic, stearic and oleic) increased (P C-16 > C-14) membrane-bound urokinase plasminogen activator (u-PA) and u-PA free binding sites in cell membranes of activated ovine blood monocytes/macrophages, but only the C-18 fatty acids (stearic, oleic) were effective towards blood neutrophils. The C-18 fatty acids up-regulated (P<005) the gene expression of u-PA, u-PA receptor, intercellular adhesion molecule 1 and inducible NO synthase (in monocytes) but not that of cyclo-oxygenase-2, integrin a X and plasminogen activator inhibitor types 1 and 2 by ovine phagocytes. SPH blocked completely or partially all C-18 fatty acid-induced changes in the expression of various pro-inflammatory genes. In conclusion, fatty acids selectively 'activate' ovine phagocytes, suggesting that these cells 'sense' metabolic signals derived from adipocytes. Soya protein peptides inhibit all changes in gene expression induced by fatty acids in ovine phagocytes in vitro. This constitutes a novel mechanism of action

Oxidative status and incidence of mastitis relative to blood &#945;-tocopherol concentrations in the postpartum period in dairy cows

Vitamin E supplementation, when combined with high blood \u3b1-tocopherol (>6.25. \u3bcg/mL) at dry off, has been reported to unexpectedly increased the risk for clinical mastitis in dairy cows. Furthermore, higher levels of oxidative stress in the postpartum period were related to higher risk of mastitis. The objective of the present study was to determine the relationship between various serum biomarkers of oxidative status, incidence of mastitis, and blood \u3b1-tocopherol concentrations at dry off and at calving. A total of 146 dairy cows from a commercial farm were used in an observational field study. All cows were supplemented with 3,000 and 50. IU/cow per day of all-. rac-\u3b1-tocopherol during the dry period and lactation, respectively. Blood samples were collected at dry off and at calving. Serum was analyzed for \u3b1-tocopherol, levels of reactive oxygen metabolites (ROM), thiol groups (SH), and ferric-reducing ability. Three \u3b1-tocopherol groups at calving were created: high (>3. \u3bcg/mL), medium (2-3. \u3bcg/mL), and low (6.25. \u3bcg/mL), medium (4.25-6.25. \u3bcg/mL), and low (<4.25. \u3bcg/mL). All cases of clinical mastitis that occurred during the dry period and the entire subsequent lactation were verified by a veterinarian. No differences were observed in the incidence of mastitis between the 3 \u3b1-tocopherol groups based on the serum levels at dry off. Incidence of mastitis was 4 times lower in the high and medium groups when compared with the corresponding value for the low-\u3b1-tocopherol group based on the serum levels at calving. Lower levels of ROM and SH at dry off and at calving were found in the group of cows with the highest \u3b1-tocopherol values at dry off when compared with the corresponding values in the low-\u3b1-tocopherol group. The ROM values at dry off but not at calving were lower in the group of cows with the highest \u3b1-tocopherol values at calving when compared with the corresponding values in the low-\u3b1-tocopherol group. No differences were observed in ferric-reducing ability values between the 3 \u3b1-tocopherol groups at dry off or calving. No differences were observed in all biomarkers of oxidative status between healthy cows and those with mastitis. Thus, blood \u3b1-tocopherol is inversely related to certain biomarkers of oxidative stress in the postpartum period and incidence of mastitis. However, reduction in the incidence of mastitis is not mediated through a reduction in the levels of various biomarkers of oxidative stress

Cloning of the 5' regulatory regions and functional characterization of the core promoters of ovine PLAU (u-PA) and SERPIN1 (PAI-1)

The activation of plasminogen plays a crucial role in various extracellular proteolytic events (fibrinolysis, cell migration, ovulation and involution of the mammary gland). In the present study we describe the isolation of the 5' proximal and distal promoter regions of ovine PLAU (urokinase plasminogen activator, u-PA) and SERPIN1 (plasminogen activator inhibitor 1, PAI-1) genes for the first time in ruminants. Analysis of the 5.645 kb 5'-flanking region of u-PA revealed a putative TATA-less promoter. In contrast the isolated 2.787 kb 5'-flanking region of PAI-1 included a TATA-box. It should be noted that both genes lack the initiator motif around the transcription start site. The two genes share a number of transcription factor binding sites, namely Nuclear Factor-kappa B, Stimulating Protein 1 and Activating protein 1, suggesting co-expression of the two genes. Moreover, additional, not shared, transcription factor binding sites were identified in u-PA and PAI-1. More important of these are the cis-regulatory elements for plasminogen activator inhibitor 2 located in the distal promoter region of u-PA, suggesting an involvement of the other specific inhibitor in the regulation of ovine u-PA gene expression, and the three stress response elements sites present in the proximal and distal promoter of PAI-1. Different genomic fragments of the two 5' flanking regions were directionally cloned into a suitable reporter vector upstream of a promoter-less luciferase gene. Transient transfection into bovine mammary epithelial (BME-UV) cells demonstrated that the regions of - 384/+27 and - 382/+22 for the u-PA and PAI-1genes, respectively, potentially function as core promoter regions

Cloning and functional characterization of the ovine Hormone Sensitive Lipase (HSL) full-length cDNAs: An integrated approach

Hormone Sensitive Lipase (HSL) is a highly regulated enzyme that mediates lipolysis in adipocytes. HSL enzymatic activity is increased by adrenergic agonists, such as catecholamines and glucagons, which induce cyclic AMP (cAMP) intracellular production, subsequently followed by the activation of Protein Kinase A (PKA) and its downstream signalling cascade reactions. Since HSL constitutes the key enzyme in the regulation of lipid stores and the only enzyme being subjected to hormonal regulation [in terms of the recently identified Adipose Triglyceride Lipase (ATGL)], the ovine Hormone Sensitive Lipase (ovHSL) full-length cDNA clones were isolated, using a Polymerase Chain Reaction-based (PCR) strategy. The two isolated isoforms ovHSL-A and ovHSL-B contain two highly homologous Open Reading Frame (ORF) regions of 2.089 Kb and 2.086 Kb, respectively, the latter having been missed the 688th triplet coding for glutamine (ΔQ688). The putative 695 and 694 amino acid respective sequences bear strong homologies with other HSL protein family members. Southern blotting analysis revealed that HSL is represented as a single copy gene in the ovine genome, while Reverse Transcription-PCR (RT-PCR) approaches unambiguously dictated its variable transcriptional expression profile in the different tissues examined. Interestingly, as undoubtedly corroborated by both RT-PCR and Western blotting analysis, ovHSL gene expression is notably enhanced in the adipose tissue during the fasting period, when lipolysis is highly increased in ruminant species. Based on the crystal structure of an Archaeoglobus fulgidus enzyme, a three-dimensional (3D) molecular model of the ovHSL putative catalytic domain was constructed, thus providing an inchoative insight into understanding the enzymatic activity and functional regulation mechanisms of the ruminant HSL gene product(s). © 2008 Elsevier B.V. All rights reserved