Characterization of Inactive and Stress-Induced Active Forms of the Transcription Factor HSF1: An analysis at the cellular level

In mammalian cells, the heat shock response is mediated by the heat shock transcription fact or HSF1 that forms active, DNA- binding homotrimers during temperature stress. The subcellular localization of the inactive form of HSF1 has been of gr eat interest for the potential implications in signaling in this system. I have detected the inactive form of HSF1 mostly in the nucleus of diverse mammalian cell lines. However , I have found that HSF1 is not confined to the nucleus , but continuously shuttles between the nucleus and the cytoplasm at a minimum rate of 1 molecule sec-1. A possible link of shuttling with the functional state of HSF1 is suggested by the observation that the shuttling cycle is discontinued during mild heat stress and resumes promptly during stress relaxation . A similar block of nuclear export is observed for deregulated mutants of HSF1 that trimerize at 37 °C, suggesting that the trimerization step inhibits an export activity. By mutational analysis I showed that HSF1 contains an unusual bipartite nuclear localization signal. Ongoing experiments are defining sequence requirements for nuclear export, most probably on a pathway distinct from Exportin-1 as judged from the refractoriness of nuclear export of HSF1 to Leptomycin B. I discuss a possible role of shuttling in compartment specific modifications of HSF1 or associations with co-factors

Quantum effects of conscious attention on the collapse of electromagnetic wave function

Uvod: Kvantni double-slit eksperiment je efekat posmatrača na klasičnu Jungovu difrakciju. Kolaps talasne funkcije podrazumeva redukciju superpozicije stanja funkcije elektromagnetnog talasa na jedno stanje dejstvom okruženja. Double-slit eksperiment se smatra za jedan od najlepših eksperimenata ikada izvedenih. Cilj: Dokazati kolaps talasne funkcije u interaktivnom polju fokusirane pažnje merenjem određenih kvantnih parametara, kao što su: korpuskularnost, kontrast, količnik intenziteta električnog polja, intenzitet interferencije na grafiku spektralne snage, kvantna povezanost, Hajzenbergova neodređenost. Materijal i metode: Eksperiment se sastoji od fizičke i fiziološke postavke sistema. U fizičkom delu sistema vršila se registracija zavisnih varijabli korišćenjem linearne ccd kamere, dok se u fiziološkom sistemu obavljala registracija nezavisnih varijabli radi kvantifikacije stanja svesne pažnje. Grupe su podeljene na kontrolnu grupu i tri eksperimentalne grupe: relaks grupa, grupa sa subjektima u stanju fokusirane pažnje i grupa sa subjektima u proceduri registracije kognitivnog potencijala P300, koja je dalje podeljena na tri podgrupe: E100, E300 i E1000 prema vremenu registracije zavisnih varijabli. U eksperimentu je učestvovalo 26 subjekata, sva snimanja su vršena tokom perioda od 45 sekundi tokom 10 nezavisnih studija odnosno sesija. Rezultati: Sa izuzetkom grupe sa subjektima u stanju relaksirane pažnje, u svim ostalim grupama dokazano je signifikantno povećanje korpuskularnih osobina elektromagnetnog talasa uz konkomitantno smanjenje interferencije i Hajzenbergove neodređenosti, kao i porast kvantne zapletenosti između fizičkog i fiziološkog kvantnog sistema u poređenju sa rezultatima u kontrolnoj grupi. Fitovanjem relacije za jačinu električnog polja na pukotinama difrakcione rešetke pokazana je značajna promena u odnosu na kontrolnu grupu u svim eksperimentalnim grupama sa izuzetkom relaks grupe. Kvantni parametri bili su statistički signifikantno izmenjeni i tokom prvih 100 milisekundi u proceduri registracije kognitivnog potencijala P300 dokazujući backward time referral efekat. Dokazano je i dejstvo P300 potencijala na vrednosti Bomove fazne razlike. Takođe, u grupi učesnika u stanju fokusirane pažnje najveća promena kvantnih parametara registrovana je tokom prve tri sekunde od dobijanja vokalne komande za početak koncentrisanja pažnje. Nezavisne varijable pokazale su porast entropije i porast determinističkog haosa. Zaključak: Dejstvom fokusirane pažnje nastaje kolaps talasne funkcije, dok značajna kvantna povezanost ukazuje na kuplung fotona, kao i na spregu fizičkog i neuronskog kvantnog sistema. Dokazana je i intervencija budućnosti na događaje u prošlosti. Promene vrednosti zavisnih varijabli reflektuju promene u neuronskom kvantnom sistemu, tako da se optički sistem može koristiti za detekciju suptilnih izmena u svesnoj pažnji.Introduction: A quantum double-slit experiment is an observer effect on classical Young diffraction. Wave function collapse occurs when a wave function in a superposition of several eigenstates reduces to a single eigenstate due to interaction with the external world. The doubleslit experiment is considered one of the most beautiful experiments ever performed. Aim: To prove the collapse of a wave function in an interactive field of focused conscious attention certain quantum parameters were measured, such as: corpuscularity, visibility, the ratio of the electric field intensity, magnitude of interference on power spectral density graph, quantum entanglement, Heisenberg uncertainty. Material and methods: The experimental procedure was divided into the physical and physiological set-up. In the physical part of the system, registration of dependent variables was performed by using a linear ccd camera, while in the physiological system, registration of independent variables was performed to quantify the state of conscious attention. The groups were divided into a control group and three experimental groups: relaxed attention group, concentrated attention group, and group for evaluating cognitive evoked potential effect on the double-slit system, which was divided into three subgroups according to time point at which data was recorded-Е100, Е300 and Е1000. Twenty-six subjects took part in the experiment, which underwent every second recording of diffraction pattern during 10 sessions, each of 45 seconds duration. Results: With the exception of the group with subjects in the state of relaxed attention, in all other groups there was a significant increase in corpuscular properties of the electromagnetic wave with a concomitant decrease in interference and Heisenberg uncertainty, as well as an increase in quantum entanglement between the physical and physiological quantum system compared with the results in the control group. The electric field ratio, which was calculated by fitting a basic equation for double slit diffraction, with exception for the relax group, showed a significant change in all experimental groups compare to control group. The quantum parameters were statistically significantly altered during the first 100 milliseconds in the cognitive evoked potential P300 registration procedure, proving a backward time referral effect. The effect of P300 potential on the values of the Bohm phase difference has also been demonstrated. Also, in concentrated attention group, the largest change in quantum parameters was registered during the first three seconds after participants started to concentrate their attention. Independent variables showed an increase in entropy as well as increase in pattern behavior known as deterministic chaos. Conclusions: This study confirmed that focused attention causes the collapse of the wave function, while significant quantum entanglement indicates photon coupling, as well as the entanglement of the physical and brain quantum systems. Backward time referral effect has also been proven. Changes in the values of dependent variables reflect changes in the brain quantum system, so that the optical system can be used to detect subtle changes in conscious attention

The influence of heat stress on metabolic status of cows

It is considered that high air temperature and humidity during the summer are the main factors which adversely affect both the health and production-reproductive performance of high yielding dairy cows. The resulting heath stress leads to a series of changes in endocrine regulation of homeostasis. The changes in hormonal status reflect in some way to the indicators of metabolic status of the cows. The objective of this work was to investigate the influence of heat stress on metabolic status of cows. The experiment was carried out on 20 cows of Holstein-Friesian breed during the summer, in the period from 18th to 45th day of lactation. During the performance of the experiment, the value of heat index (THI) was determined hourly and then the value of average morning (from 10 pm the previous day to 9 am the current day), afternoon (from 10 am to 9 pm the current day) and all-day THI was calculated. Blood sampling was carried out on the 1st, 2nd, 8th, 11th, 14th, 18th, 25th, 29th and 37th day of the experiment, in the morning and the afternoon. On the basis of hourly THI values, whole experimental period was divided into three periods: period A during which the cows were exposed to a extreme high heat stress (THI≥78) at least 7 hours in 24 hours; period B during which the cows were exposed to a moderate heat stress (72≥THI≤78) at least 7 hours in 24 hours; period C during which the cows were not exposed to a heat stress (THI≤72) in 24 hours. The average daily THI in period A (73,25±0,89) was significantly higher (p<0,01, individually) in regard to period B (71,45±0,96) and period C (65,41±2,09). THI was significantly higher in the period B than in the period C (p<0,01). Significantly lower blood glucose value (p<0,05) during the afternoon period in the cows exposed to the extreme heat stress (3,02±0,31 mmol/L) in regard to the morning period (3,14±0,41 mmol/L) points to the fact that in such conditions, metabolism redirects to use of glucose as an energy source because in that way less thermal energy is produced than during decomposition of fatty acids. Concentration of most important metabolic profile parameters in blood (cholesterol, triglycerides, total proteins, albumin, urea, total bilirubin, calcium, inorganic phosphorus as well as AST and ALT activity) did not vary significantly under the influence of heat stress. The exception was ionic calcium concentration which, under the conditions of extreme heat stress, was on the lower limit of physiological values (1.17±0.16 mmol/L)

Relationship between the indexes of insulin resistance and metabolic status in dairy cows during early lactation

Insulin resistance is a phenomenon which accompanies the ongoing metabolic adaptation in cows during early lactation. The aim of our study was to determine the linear correlations of HOMA (Homeostatic Model Assessment), QUICKI (Quantitative Insulin Sensitivity Check Index) and RQUICKI (Revised Quantitative Insulin Sensitivity Check Index) indexes of insulin resistance with the metabolic status of cows (concentration of hormones, metabolites and body condition score). The experiment included 40 Holstein-Frisian cows in the first week after calving. Indexes of insulin resistance valued: 18.68 +/- 5.43 (HOMA), 0.39 +/- 0.06 (QUICKI) and 0.45 +/- 0.06 (RQUICKI). Linear correlations were examined by testing the coefficient of correlation (r), determination (r(2),%) and regression parameter beta (b) in linear equation. A negative correlation was found between HOMA and IGF-I (insulin growth factor I) (r=-0.51, r(2)=25.0, b=-1.1257, p<0.01). HOMA showed a positive correlation with BHB (beta-hidroxybutyrate) (r=0.48, r(2)=23.2, b=0.0234, p<0.01). A positive correlation was found between QUICKI and IGF-I (r=0.30, r(2)=10.0 b=46.7900, p<0.05) and cholesterol (r=0.44, r(2)=18.3, b=1.9021, p<0.01). In contrast, QUICKI and BHB (r=0.51, r(2)=27.1, b=-1.7241, p<0.01), just like QUICKI and BCS (r=0.46, r(2)=20.9, b=-2.424, p<0.01), showed a negative correlation. RQUICKI showed positive correlations with IGF-I (r=0.48, r(2)=22.8, b=28.1230, p<0.01), T4 (r=0.47, r(2)=22.1, b=87.142, p<0.01) and triglycerides (r=0.36, r(2)=13, b=0.0407, p<0.05) but negative correlations with cortisol (r=-0.36, r(2)=13.0, b=-9.0332, p<0.05), STH (somatotropic hormone) (r=-0.42, r(2)=17.3, b=-5.4976, p<0.01), BHB (r=-0.62, r(2)=38.3, b=-1.1872, p<0.01), total bilirubin (r=0.58, r(2)=33.7, b=-7.131, p<0.01) and BCS (body condition score) (r=-0.6, r(2)=36.4, b=1.8347, p<0.01). In conclusion, indexes of insulin resistance may be used to evaluate the metabolic status of cows in early lactation. RQUICKI might be the most appropriate predictor of metabolic status due to its linear relationship with most of the parameters included in homeorhetic process

Effects of buffering mineral mixtures on milk yield, milk composition, rumen pH and some blood biochemical parameters in heat stressed dairy cows

The objective of the work was to investigate the influence of partial substitution of magnesium oxide with natural bentonite in feed mixtures used in feeding of cows during their exposure to heat stress. The investigation lasted 30 days and was carried out during last ten days of may and first twenty days of June when average air temperature in stables was 36.6±2.5oC. In the experiment there were included 30 cows of Holstein breed in first phase of lactation, which were divided into two groups of 15 cows: control (C) and experimental (E). Group C was fed with experimental mineral mixture that contained 60% of magnesium oxide during the whole investigation period. Group E was fed with experimental mineral mixture that contained 40% of magnesium oxide as well as 20% of natural bentonite. Remaining ingredients in both control and experimental mineral mixtures were the same and also contained 20% of sodium bicarbonate and 20% of zeolite in the same quantities. The control and experimental mineral mixtures were mixed into complete feed mixture (18% UP) in the amount of 1%. At tne end of the investigation period, on the 30th day, there were taken samples of rumens contents for determining pH, and after that blood samples, in which, after the separation of blood serum, were determined glucose concentration, total proteins, albumin, globulin, urea, HDL cholesterol, LDL cholesterol, total bilirubin, calcium and phosphorus, as well as the activity of ALT and AST. By computation there was calculated the ratio between albumin and globulin, ALT and AST, and the ratio between calcium and phosphorus. Daily allowance and milk chemical composition ( percentage of fat, proteins and dry substance) were determined at the end of the investigation period, that is on the 30th day of lactation, for each cow individually. Partial substitution of magnesium oxide with bentonite influenced milk production increase, but it was statistically insignificant. Besides that, in E group of cows, percentage of fat and dry substance in milk was significantly increased (p<0.05 and 0.01 respectively), while percentage of protein increase was insignificant. The substitution of magnesium oxide with bentonite had no impact on the values of examined parameters of metabolic profile, but it led to statistically significant increase of rumen contents pH values (p<0.05). From the obtained results it can be concluded that substitution of magnesium oxide with bentonite in feed mixtures that are used for feeding cows during summer period, can prevent rumen acidosis, which high yielding cows incline to under the conditions of elevated external temperatures. Besides that, this kind of substitution leads to improvement of milk composition, especially when percentage of fat and dry substance in milk is concerned. [Projekat Ministarstva nauke Republike Srbije, br. TR 31003 i br. TR 34013

Tribological Aspects of the Process of Winding the Steel Rope Around the Winch Drum

Proper winding of the steel rope around the winch drum is great importance, mostly for: prolonging the service life of the rope, reduction of deformations of the body and the sides of the drum if the winding of the rope is multilayered, increasing of the safety factors, easier unwinding of the rope while lowering the load, even running of the drive unit, etc. The focus of this paper is on the analysis of the friction which occurs in the process of winding and unwinding the rope around the winch drum. Friction force is in its highest intensity when the rope passes from one layer to another, if the winding of the rope is multilayered. As the result of the research, certain mechanisms of winding of the rope from the aspects of the friction force were obtained, and the effects of the forces on the sides of the drum were analyzed

Uticaj polukružnih veza elemenata paletnih regala na globalnu analizu konstrukcije

Predmet proučavanja ove disertacije su veze koje se ostvaruju između elemenata čelične konstrukcije paletnih regala. Vertikalni ramovi i horizontalne grede, najčešće proizvedeni od tankozidnih hladno oblikovanih profila formiraju prostorni ramni sistem paletnih regala. Ramovi leže u vertikalnoj ravni poprečno na pravac glavnog hodnika regala. Sastoje se od dva perforirana stuba međusobno povezana sistemom dijagonalnih i/ili horizontalnih ukrućenja zavarenih ili zavrtnjevima vezanim za stubove. Ovaj sistem ukrućenja obezbeđuje stabilnost regala u poprečnom pravcu. Grede povezuju susedne ramove i leže u podužnom pravcu paralelno glavnom hodniku regala. Gredni konektori su delovi grede zavareni ili na drugi način formirani kao njen sastavni deo i imaju posebne delove koji upadaju u otvore ili proreze na stubu. Podužna stabilnost regala se prvenstveno obezbeđuje krutošću veze greda-stub. U praksi postoje različiti tipovi i dizajni ovih veza, karakteristični za različite proizvođače regala. U disertaciji je obrađena klasifikacija i modeliranje veza u skladu sa Evrokodom 3. Kako je nemoguće da se razvije opšti analitički model proračuna ovih veza, trenutno jedini način da se odredi karakteristika takve veze je eksperiment. U disertaciji su prikazani postupak i rezultati ispitivanja veze greda-stub po proceduri definisanoj u FEM propisima u cilju određivanja krive moment-rotacija (M- krive). Uticaj veza na globalno ponašanje konstrukcije regala uvodi se pomoću M- krive, odnosno konstrukcijskih svojstava veze koje se iz nje određuju: krutost, otpornost i rotacioni kapacitet. Kako bi se izbegao veliki broj skupih ispitivanja u cilju određivanja M- krive za različite tipove veza kojih u praksi može biti jako puno, u okviru ove disertacije je razvijen numerički model za simulaciju eksperimenta primenom metode konačnih elemenata. Nakon verifikacije modela raspoloživim eksperimentalnim rezultatima, isti se može primeniti na najrazličitije kombinacije veze greda-stub. Pouzdanim određivanjem konstrukcijskih svojstava veze primenom razvijenog modela može se vršiti globalna analiza ponašanja konstrukcije kao i proračun pojedinih elemenata po postupcima definisanim u FEM propisima.The case study of this dissertation are the connections that are established between elements of the steel structure of pallet racks. The vertical frames and horizontal beams, usually made of thin-walled cold-formed profiles form a spatial frame structure of pallet racking system. Upright frames lie in the vertical plane, in the cross aisle direction, normal to the main aisle of the rack. They consist of two perforated uprights linked together by a system of diagonal and/оr horizontal bracing welded or bolted to the uprights. This bracing system provide rack stability in cross-aisle direction. Beams connecting adjacent frames and lying in the horizontal direction parallel to the main aisle. Beam-end connectors are welded to or otherwise formed as an integral part of the beams, which has special devices which engage in holes or slots in the upright. The down-aisle stability primary is provided by the stiffness of the semi-rigid joints between uprights and beams. In practice, there are different types and designs of these connections, which characterize the different racks manufacturers. In dissertation classification and modelling of connections has been treated according to Eurocode 3. As it is impossible to develop a general analytical model for calculating these connections, currently the only way to determine the properties of such connections is an experiment. The dissertation presents the test procedure and results of beam-to-column connections according to the procedure defined in the FEM recommendations in order to determine the moment-rotation curve (M- curve). Influence of connections to the global structural racks behavior introduced with M- curve and its structural properties: stiffness, bending strength and rotational capacity. In order to avoid a large number of expensive tests with aim to determine the M- curves for different types of connection which in practice may be very much, part of this dissertation developed a numerical model to simulate the experiment using the finite element method. After verification of the model to the available experimental results, it can be applied to various combinations of beam-to-column connections. Reliable determination of structural properties of the connection using the developed model can be made a global analysis of the structural behavior and the calculation of elements according to the procedures defined in the FEM recommendations

The biological significance of differences in cows and sows colostrum and milk composition

The objective of this work was to compare the composition of colostrum and milk of cows and sows (content of dry matter, protein, milk fat and lactose, concentration of IGF-I and insulin) in samples taken on the first, second, third and seventh day after parturition, and then based on the differences in composition to determine a biological significance of nutrition of newborn during the earliest stages of their life. The investigation inluded 14 cows of Holstein breed and 14 sows of Landrace breed. The content of dry matter and the concentration of proteins in both colostrum and milk samples were statistically significantly higher in regard to sows mammary glands secretion, taken on the first day after the parturition (p<0,01 and p<0,001, individually), but their decrease in mammary glands secretion was more pronounced in the cows than the sows, during the first seven days. The concentration IGF-I was statistically significantly higher in the cows colostrum and milk in regard to the sows during the whole investigation period, while the concentration of insulin was significantly higher in the sows in regard to the cows during the same period. The concentrations of milk fat and lactose in cows milk samples were significantly lower in regard to the sows in all period of the study. On the basis of the obtained results, it can be concluded that there are significant differences in the composition of milk and colostrum of both the investigated animal species. The differences are probably the result of evolutionary adaptation of mammal gland function to nutrition, energy and protection requirements of these young animals in their early postnatal life. [Projekat Ministarstva nauke Republike Srbije, br. III 46002

Puerperalna hemoglobinurija - Haemoglobinuria Puerperalis

Puerperal haemoglobinuria is a sporadic disease that appears in cows before the fourth week after calving and is manifested as haemoglobinuria and haemolytic anaemia. Haemoglobinuria puerperalis was diagnozed at a private mini cattle farm on the grounds of anamnestic data and the results of clinical examinations, and it was confirmed with the results of laboratory examinations of blood samples. In the course of the disease, characteristic symptoms developed, there was clearly visible paleness of the mucosa, slightly expressed ichtherus of the eyelid ridge skin, and haemoglobinuria. Having in mind the very expressed oligocytemic hypochromic anaemia, therapy was directed at the primary disease and a blood transfusion was administered.Puerperalna hemoglobinurija je sporadično oboljenje krava koje se pojavljuje do četvrte nedelje posle teljenja, a manifestuje se hemoglobinurijom i hemolitičkom anemijom. Na privatnoj mini-farmi goveda na osnovu anamnestičkih podataka i rezultata kliničkog pregleda postavljena je dijagnoza - Haemoglobinuria puerperalis, potvrđena rezultatima laboratorijskog ispitivanja uzorka krvi. U toku bolesti je došlo do razvoja karakterističnih simptoma - jasno uočljivo bledilo vidljivih sluznica, blago izražen ikterus ruba kože kapaka i hemoglobinurija. S obzirom na veoma izraženu oligocitemičnu hipohromnu anemiju, terapija je bila usmerena prema primarnoj bolesti i primenjena je transfuzija krvi

Thyroid Hormones Concentrations during the Mid-Dry Period: An Early Indicator of Fatty Liver in Holstein-Friesian Dairy Cows

Relationship between postpartal fatty liver and thyroid gland activity during the peripartal and mid dry periods was studied. Twenty one dry cows were chosen. Blood samples were obtained on days −30, −2, and +12 related to calving and analized for thyroxine (T4) and triiodothyronine (T3). A T3/T4 ratio was calculated. Liver tissue samples were taken 12 d after calving and tested for the lipid content. Cows were divided into three groups: mild (<20% fat), moderate (20 to 30%), or severe fatty liver (>30%). Cows, that were affected with severe fatty liver, were hypothyroid prior to development of the condition due to lower T4 concentrations, and had significantly lower concentration of T3 and higher T3/T4 ratios than cows with mild and moderate fatty liver. Thus, hypothyroid state during mid-dry period may be an early indicator of postpartal fatty liver and may provoke T3/T4 ratio increase in this group of cows